ライフサイエンスコーパス: nicking

1 y in apoptotic cells, around the time of DNA nicking.

2 orms and converts them to open forms without nicking.

3 was the only ICEBs1 gene product needed for nicking.

4 dest effect on stimulation of MutH-catalyzed nicking.

5 rated from its complementary strand prior to nicking.

6 biting religation and to a lesser extent DNA nicking.

7 sation and collapse, and single-stranded DNA nicking.

8 group is important only for steps preceding nicking.

9 tion amplification followed by site-specific nicking.

10 educe the catalytic rate constant k(cat) for nicking.

11 purified RAG enzymatic activity at both the nicking (2- to 5-fold) and hairpinning (3- to 11-fold) s

12 synthesis and ligand-dependent TOP1-mediated nicking-a strategy exerting quantitative effects on eRNA

13 The three genes code for a nicking accessory protein, a relaxase, and a VirD4-like

14 Glycosylase nicking activities were measured on both thymine glycol-

15 gent, displaying significant supercoiled DNA-nicking activity at concentrations as low as 1 microM.

16 to the H-N-H endonuclease family and possess nicking activity in vitro.

17 ZFNickases show robust strand-specific nicking activity in vitro.

18 ntal evidence, we propose a model where RepC nicking activity is passive and dependent upon the super

19 The nicking activity is similar between the double-stranded

20 -modified DNA in Ni2+ buffer even though the nicking activity is sub-optimal compared to the activity

21 The nicking activity of AAV-2's Rep68 or Rep78 proteins is e

22 The AP nicking activity of the C136R variant was significantly

23 The DNA nicking activity of the wild-type enzyme is still activa

24 n sites on dsDNA generates in some cases the nicking activity of these DNA cutters.

25 Furthermore, we show that the DNA nicking activity of TOP1 is a prerequisite for robust eR

26 nt degree of DNA protection against cleaving/nicking activity of various isoschizomeric endonucleases

27 and D252N) retained near wild type levels of nicking activity on abasic (AP) site-containing DNA, G83

28 safe harbor" locus revealed similar in vitro nicking activity, a marked reduction of indels character

29 e first time, we show Rep protein off-target nicking activity, highlighting the importance of the nic

30 Mutation at mcb2 abolished detectable nicking activity, suggesting that binding of this site b

31 or head-to-tail repeat only supported a DNA nicking activity.

32 th significant strand- and sequence-specific nicking activity.

33 It investigates the phenomenon that nicking, although required to open a single-stranded reg

34 reover, it performed primary "polymerization-nicking" amplification and mediate secondary HRCA.

35 replication origin, which contains both NS1 nicking and binding sites, to a 46-nucleotide sequence i

36 -deoxyguanosine (N2-ethyl-dG), on human Top1 nicking and closing activities.

37 developed two biochemical methods; i.e., DNA-nicking and DNA-gyrase methods to examine whether certai

38 , the physiologically relevant metal ion, in nicking and double-strand cleavage of both RSS DNAs to p

39 ion in RAG1 had an inhibitory effect on both nicking and hairpin stages that could be rescued by abas

40 Kinetic analysis revealed that first-strand nicking and integration are rapid.

41 d signs of retinal microvascular lesions (AV nicking and microaneurysms/hemorrhages) are more likely

42 The cloning and expression of the CviPII DNA nicking and modification system encoded by chlorella vir

43 Following nicking and powered by ATP hydrolysis, the concerted act

44 plasmid-encoded protein MobA, which has both nicking and priming activities on single-stranded DNA, i

45 of the probe residue retards the kinetics of nicking and reduces base flipping by 50%.

46 Nicking and reduction render both CT and the closely rel

47 axase enzyme initiates and terminates CPT by nicking and religating the transferred plasmid in a sequ

48 agnetic tweezers to directly measure the DNA nicking and religation activities of RepC, the replicati

49 f this cruciform and the implication for its nicking and religation functions is unclear.

50 ues (Y18,19,26,27) that are required for DNA nicking and religation were displaced up to 14 A out of

51 Top1 relieves torsional stress by nicking and resealing one DNA strand, and some Top1-depe

52 vector is then linearized by digestion with nicking and restriction endonucleases.

53 The spacer sequences between the nicking and restriction sites are tailored to create ss

54 The nicking and reverse transcription steps in the reaction

55 echanism of botulinum neurotoxin type E upon nicking and subsequent reduction of disulfide bond.

56 gnal sequence (RSS)-conserved regions before nicking and synapsis.

57 DNA cleavage by affecting either or both the nicking and the closing activities of Top1 depending on

58 rved tyrosine, Tyr127, are essential for DNA nicking and the formation of a covalent intermediate bet

59 bination signal sequences through sequential nicking and transesterification reactions to yield blunt

60 ise from DNA regions expected to undergo DNA nicking and/or double-strand breaks.

61 Metnase possesses a unique DNA nicking and/or endonuclease activity that mediates cleav

62 ange within the nucleosome to facilitate DNA nicking and/or fragmentation.

63 focal arteriolar narrowing and arteriovenous nicking) and retinopathy lesions (retinal blot hemorrhag

64 tivities of Rep68, including DNA binding and nicking, and compromises viral DNA replication and trans

65 cal arteriolar narrowing, arteriovenous (AV) nicking, and microaneurysms/hemorrhages were evaluated o

66 y, focal arteriolar narrowing, arteriovenous nicking, and the arteriovenous ratio--the last based upo

67 ochemical activities, including DNA binding, nicking, and unwinding.

68 hat residues responsible for NS1 binding and nicking are within the origin-binding domain.

69 t also may affect the specificity of hairpin nicking, as well as the efficiency of joining open codin

70 was determined in plasmid using the plasmid-nicking assay and in cells by either conventional pulsed

71 oxyribose oxidation as measured by a plasmid nicking assay.

72 their substrate identity in a MobA-specific nicking assay.

73 aks and abasic sites determined by a plasmid nicking assay; the abasic sites comprised less than 10%

74 tive double-stranded substrates in gel-based nicking assays.

75 RNaseH2 must be efficient at recognizing and nicking at embedded ribonucleotides to ensure genome int

76 levant, nonamer-independent mechanism of RAG nicking at mcr, which may be important for generation of

77 icK likely mediates conjugation of ICEBs1 by nicking at oriT and facilitating the translocation of a

78 phi29 DNA was modified in vitro by nicking at random sites with DNase I, or at specific sit

79 These results support the hypothesis that nicking at the conjugal origin initiates the recombinati

80 Further, we demonstrate that RAG binding and nicking at the mcr are independent of nonamer, whereas a

81 tiate transfer by catalyzing strand-specific nicking at the pCF10 origin of transfer sequence (oriT).

82 Arteriovenous nickings (AVNs) in the retina are the cause of retinal v

83 ized CRISPR/Cas tools can be used for offset nicking-based mutagenesis.

84 nce arises from preferential endoproteolytic nicking between stable segments followed by loading of f

85 Relative to a single nick, nicking both strands enhances HR, consistent with a DSB

86 We found that the overall rate of nicking, but not hairpinning, is critical for the discri

87 predicted sites of metal cation binding, but nicking by chymase does not alter binding of Cu2+ or Zn2

88 g of this site by MobC is a prerequisite for nicking by MobA.

89 7 bp sequence, sra, which was essential for nicking by MobA.

90 Nicking by RepC occurred only in negatively supercoiled

91 The combined continuous repetitive action of nicking by the endonuclease and strand-displacement synt

92 In the case of staphylococcal plasmid pC221, nicking by the plasmid-specific MobA relaxase is facilit

93 be a prerequisite step for the site-specific nicking by the V(D)J recombinase.

94 de strand, and Cas9 nickases used for paired nicking can also tolerate bulges in one of the guide str

95 We demonstrate that using paired nicking can reduce off-target activity by 50- to 1,500-f

96 smid pCU1 and determined its DNA binding and nicking capabilities.

97 PleI isoschizomer, does not exhibit any DNA nicking/cleavage activity, being completely blocked by t

98 purified recombinant Top1, we show that Top1 nicking-closing activity remains unaffected in N2-ethyl-

99 In addition to its classic DNA nicking-closing functions, Top1 plays critical nonclassi

100 Paradoxically, the MMR-nicking complex Pms2/Mlh1 is apparently dispensable for

101 The rate of nicking correlates with the iron content of the ferritin

102 We used the double-nicking CRISPR/Cas9 system to conduct site-specific muta

103 primers released from above "polymerization-nicking" cycles were separated out to trigger the subseq

104 SP82 introns, the Bastille intron encodes a nicking DNA endonuclease of the H-N-H family, I-BasI, wi

105 I-HmuI and I-BasI are two highly similar nicking DNA endonucleases, which are each encoded by a g

106 yses four phosphoryl transfer reactions (DNA nicking, DNA hairpin formation, hairpin resolution and s

107 Arteriovenous nickings do not necessarily involve an arteriovenous com

108 The small modular HNH nicking domain can be used to generate rare NEases appli

109 A minimal HNH nicking domain of 76 amino acid residues was identified

110 The nicking domains (NDs) of E59A and 11 double mutants were

111 e replication tracks that yield the Nt.BbvCI nicking domains and the respective Mg(2+)-dependent DNAz

112 genes and replication tracks that yield the nicking domains for Nt.BbvCI and two different Mg(2+)-de

113 merase that relaxes supercoils by reversibly nicking duplex DNA.

114 DA), the T4 replisome is supplemented with a nicking endonuclease (Nb.BbvCI) and a reverse primer to

115 min by means of thermostable polymerase and nicking endonuclease activities.

116 A into a pre-modified host and screening for nicking endonuclease activity in total cell culture or c

117 tion of the sequence motif recognized by the nicking endonuclease can be established with good accura

118 to initiate the polymerization extension and nicking endonuclease cleavage reaction.

119 o-linear MTase encoding gene (cviPIIM) and a nicking endonuclease encoding gene (cviPIINt) separated

120 at specific sequence motifs by the action of nicking endonuclease followed by the incorporation of dy

121 g endonuclease, then using sequence-specific nicking endonuclease N.AlwI to introduce single nicks fo

122 er, purified SCD protein acts a non-specific nicking endonuclease on normal linear DNA.

123 gle- or double-stranded DNA sequences called nicking endonuclease signal amplification (NESA).

124 tains a cassette with two inversely oriented nicking endonuclease sites separated by restriction endo

125 ucleotides can be linearly amplified using a nicking endonuclease with 7-bp recognition sequence and

126 s generated using sequence-specific N.BstNBI nicking endonuclease, then using sequence-specific nicki

127 ee-arm DNA junction, subsequently initiating nicking endonuclease-assisted isothermal fluorescence si

128 this system in two formats: (i) in circular nicking endonuclease-dependent amplification (cNDA), the

129 or linear isothermal DNA amplification using nicking endonuclease-mediated strand displacement by a D

130 emonstration of intron homing initiated by a nicking endonuclease.

131 HNHEs are site-specific and strand-specific nicking endonucleases (NEase or nickase) with 3- to 7-bp

132 ation of vicinal nicks, which are created by nicking endonucleases (NEases) at specified DNA sites on

133 Naturally occurring nicking endonucleases (NEases) range from frequent cutte

134 s observe the response of supercoiled DNA to nicking endonucleases and topoisomerases.

135 st, only a limited number of strand-specific nicking endonucleases are currently available.

136 Sequence-specific nicking endonucleases are used to create the 4-nt-long g

137 ases, RNase H2 can be replaced with specific nicking endonucleases in this protocol; we term this met

138 TR include sequence-dependent restriction or nicking endonucleases or sequence independent exonucleas

139 The number of strand-specific nicking endonucleases that are currently available for l

140 Nt.BsaI and Nb.BsaI nicking endonucleases were isolated from BsaI using this

141 he lacZalpha gene with sites for two related nicking endonucleases, enabling the excision of either t

142 candidates for engineering high-specificity nicking endonucleases.

143 striction endonucleases into strand-specific nicking endonucleases.

144 By the second method, a nicking enzyme (Nt.BspQI) is added to the system, and th

145 a is a strand-specific and site-specific DNA nicking enzyme (YCG downward arrowGT or AC upward arrowC

146 Here, we describe NicE-seq (nicking enzyme assisted sequencing) for high-resolution

147 lar basis for resistance transmission by the nicking enzyme in S. aureus (NES), which is essential fo

148 ites with DNase I, or at specific sites with nicking enzyme N.BbvC IA.

149 ATP- and NAD+-dependent DNA ligases and the nicking enzyme N.BbvCIA.

150 basic version, though simple, requires that nicking enzyme recognition sequence be present in the ta

151 rporates a G-quadruplex structure flanked by nicking enzyme recognition sites.

152 introduce a scheme of signal amplification, nicking enzyme signal amplification, to increase the det

153 Cas9 can also be converted into a nicking enzyme to facilitate homology-directed repair wi

154 wo coadded components ("helper" sequence and nicking enzyme), the sensitive detection of the analyte

155 he conjugal origin, we supplied an exogenous nicking enzyme, the gene II protein (gIIp) of bacterioph

156 ndent cleavage of molecular beacons by a DNA nicking enzyme, through which one target DNA can open ma

157 io-bar-code amplification (BCA) and Nb.BbvCI nicking enzyme-assisted strand cycle for exponential sig

158 serted at user-defined sites via an improved nicking enzyme-based strategy.

159 tivities of Bst 2.0 polymerase and Nt.BstNBI nicking enzyme.

160 rge subunits behave as sequence-specific DNA nicking enzymes and only nick the bottom strand of the s

161 These DNA-nicking enzymes could prove useful for investigation of

162 The development of sequence-specific nicking enzymes like the I-AniI nickase will facilitate

163 procedure designed to isolate bottom-strand nicking enzymes yielded Nb.SapI-1 containing a critical

164 hyltransferases and related proteins such as nicking enzymes, specificity subunits and control protei

165 NA sensors and aptasensors using replication/nicking enzymes/DNAzyme machineries is described.

166 ucture is apparently recognized by cellular "nicking" enzymes.

167 p downstream from the Top1 site enhances DNA nicking especially when the 3' end of the TFO is proxima

168 requirement for cooperativity through offset nicking for genome editing or through multiple synergist

169 protein shows specificity for binding to and nicking forked DNA within single strand gaps, and collap

170 fer to the single subunit, the bottom-strand nicking forms as 'hemidimers'.

171 variants, and the applications of NEases and nicking HEases.

172 s characteristic has also resulted in useful nicking HEases.

173 F recruits PcfG to oriT to catalyze T-strand nicking, (ii) PcfF and PcfG spatially position the relax

174 -established significance of strand-specific nicking in MMR, the mechanism(s) by which MutS and MutL

175 Our laboratory previously demonstrated that nicking in the RGGG sequences occurs preferentially 5' t

176 ruses were both able to support NS1-mediated nicking in vitro, but initiation efficiency was consiste

177 ion and SET cleavage at N175, triggering DNA nicking in wild-type, but not AEP null, mice.

178 led plasmid DNA from oxidative damage (i.e., nicking) in vitro.

179 We propose the top-strand nicking is carried out by a Tth111II monomer and bottom-

180 Ferritin-dependent nicking is characterized by a kinetic lag that is not se

181 Nicking is controlled largely by the coding flank and, u

182 dent cell death in which DNA single-stranded nicking is observed instead of oligonucleosomal fragment

183 Nicking is performed by a replication-initiation protein

184 Recruitment and nicking is stimulated by the presence, but not hydrolysi

185 essed in bacteria yielded protein containing nicking-joining activity, implying that K4L is the only

186 only vaccinia virus protein required for the nicking-joining enzymatic activity.

187 Nicking leaves NS1 covalently attached to the 5' end of

188 NS1-mediated nicking liberates a base-paired 3' nucleotide to prime D

189 ndary coupled activation of a polymerization/nicking machinery and DNAzyme generation path leads to a

190 DNA one base 3' to the mismatch, as well as nicking matched DNA at low levels.

191 Here, we observed that venous nicking may be observed in the absence of physical conta

192 his strain of E. coli, and they suggest that nicking may be the only conjugal function required for a

193 on from ferritin is an important part of the nicking mechanism.

194 The current nicking model suggests that the strand containing the ni

195 Here we present nicking mutagenesis, a robust, single-day, one-pot satur

196 us segments showed a variable association of nicking, narrowing, deviation, and opacification.

197 a coli produces modified DNA, on which EndoV nicking near the base analogues initiates excision repai

198 el in which the donor backbone DNA, prior to nicking, occupies the same space that is invaded by the

199 d D-loops, which is primarily in the form of nicking, occurs to a greater extent at a temperature tha

200 ation with the presence of either retinal AV nicking (odds ratio [OR] 2.47 [95% CI 1.42-4.31]) or ret

201 It was observed that nicking of BoNT/E significantly enhances its endopeptida

202 intracellular Chi hotspot activity and that nicking of DNA by RecBCD enzyme at Chi is sufficient.

203 recognition of the origin of replication and nicking of DNA during bocavirus genome replication.

204 t not general oxidative damage or UV-induced nicking of DNA, caused H2Av phosphorylation, suggesting

205 Unusually, concentration-dependent nicking of duplex DNA appeared to require only transient

206 we show that the site-specific PNA-assisted nicking of duplex DNA can be engaged in a rolling-circle

207 ability to effect single- and double-strand nicking of duplex DNA, sequence-selective DNA cleavage,

208 ORF1p depletion and reduced the L1-mediated nicking of genomic DNA.

209 Optimal nicking of heteroduplexes for mismatch detection was ach

210 Expression of nicK leads to nicking of ICEBs1 between a GC-rich inverted repeat in o

211 hat the B12/23 rule is enforced through slow nicking of Jbeta substrates and to some extent through p

212 equence-specific recognition for initiation, nicking of one of the template DNA strands and unwinding

213 ase domain in Chi recognition, indicate that nicking of one strand at Chi is RecBCD's biologically im

214 The nicking of one strand of a DNA target by the endonucleas

215 s the F origin of transfer (oriT) to promote nicking of plasmid DNA prior to conjugative transfer.

216 and that these proteins enhance binding and nicking of Rep proteins at the origin.

217 totoxic 1,2-dialkynylimidazoles do not cause nicking of supercoiled plasmid DNA or cleavage of bovine

218 We also showed that nicking of the bottom strand by I-TevI is not affected b

219 depending on the reaction conditions: simple nicking of the Chi-containing strand at Chi or switching

220 rotation than the closed state required for nicking of the DNA.

221 The nicking of the origin of transfer (oriT) is an essential

222 Nicking of the single chain BoNT/E to the dichain form i

223 sh2-Msh3), PCNA, and RFC but did not require nicking of the substrate, followed by a second stage in

224 PA(63), generated by protease "nicking" of whole PA, is responsible for delivering the

225 re demonstrated via the marked effect of DNA nicking on histone eviction that underscores the powerfu

226 Cleavage occurs by nicking one DNA strand, followed by a one-step transeste

227 supercoils by clamping a duplex DNA segment, nicking one strand at a phosphodiester bond, covalently

228 ranscription and recombination by reversibly nicking one strand of duplex DNA and forming a covalent

229 s were designed to isolate either top-strand nicking or bottom-strand nicking variants from a randoml

230 ir of sequence-specific nuclease-induced DNA nicking or double-strand breaks (DSBs) by homology-direc

231 several step arrest mutants with defects in nicking or hairpin formation, and four RAG-1 mutants def

232 The nicking, or relaxase, activity has been localized to the

233 interact with MutS, stimulate MutH-catalyzed nicking, or stimulate the loading of UvrD.

234 s, Orf20 functions as the relaxase of Tn916, nicking oriT as the first step in conjugal DNA transfer.

235 ones were segregated to confirm a top-strand nicking phenotype for single variants Q240R, E250K, G271

236 espective recognition sequences triggers the nicking/polymerization machineries, leading to the synth

237 rs preferentially 5' to a G residue with the nicking probability decreasing from the 5' to 3'end of t

238 -triggered isothermal autonomous replication/nicking process on the modified template results in the

239 ger on the autonomous isothermal replication/nicking processes and the displacement of a Mg(2+)-depen

240 imated to be much slower than the top-strand nicking rate.

241 also a useful tool for dissecting individual nicking rates of a double-stranded cleavage.

242 with MutS and stimulated the MutH-catalyzed nicking reaction in a mismatch-dependent manner.

243 The equilibrium constant of the nicking reaction, which involves a transesterification t

244 unctional magnetic probes and polymerization nicking reactions mediated hyperbranched rolling circle

245 les, as it contains both single-stranded DNA-nicking relaxase and ATP-dependent helicase domains with

246 pendent DNAzyme sequences are implemented as nicking/replication machineries for the amplified, multi

247 5mCG-dependent nicking requires special digestion conditions in high sa

248 s human site, including identical spacer and nicking sequences.

249 Both single gRNA/WT hCas9 and double nicking set-ups were effective.

250 ividual roles for the RBE and the spacer and nicking site elements.

251 The region flanking the nicking site in AAVS1 (5'-GGCGGCGGT/TGGGGCTCG-3' [the sl

252 e that ZFNickases can stimulate HDR at their nicking site in human cells, albeit at a lower frequency

253 placed into a single-stranded form while the nicking site is also regenerated.

254 In AAV serotypes 1 through 6, the nicking site is flanked by a sequence that is predicted

255 odel suggests that the strand containing the nicking site is separated from its complementary strand

256 are outside the previously recognized GTTGG nicking site motif and include parts of the CTC motif th

257 ild-type or mutated sequences from the AAVS1 nicking site to appropriate standards, on native and den

258 f mobA, and a putative Bacteroides consensus nicking site was observed within this region.

259 otides are then incorporated adjacent to the nicking site with a DNA polymerase to label the guide RN

260 68/78), a Rep68/78 binding site (RBS), and a nicking site within AAVS1 and may also require an RBS wi

261 ases and were all downstream of the Rep68/78 nicking site within AAVS1.

262 ced levels of mutagenic NHEJ at their target nicking site.

263 V Rep binding element (RBE), a spacer, and a nicking site.

264 GGCGGT/TGGGGCTCG-3' [the slash indicates the nicking site]) lacks extensive potential for Watson-Cric

265 with Illumina primer sequences and specific nicking sites flanking the cloning site.

266 lyses confirmed the putative NS1 binding and nicking sites within the OriR.

267 ration of fluorophore-labeled nucleotides on nicking sites.

268 We found that Artemis is capable of nicking small heteroduplex structures and is even able t

269 The latter extends the range of enzymes for nicking specified strands at specific sequences, and may

270 that no greater DNA bending occurs after the nicking step in the signal complex.

271 A elements and strongly suggest that the DNA nicking step of V(D)J recombination can be rate limiting

272 arget cleavage, we further describe a double-nicking strategy using the Cas9 nickase mutant with pair

273 Recently a "double-nicking" strategy using catalytic mutant Cas9(D10A) nick

274 activity, highlighting the importance of the nicking substrate for Rep-mediated integration.

275 ses, within the context of a double-stranded nicking substrate, reduces the ability of the substrate

276 nucleic acid (PNA)-directed design of a DNA-nicking system that enables selective and quantitative c

277 most bacteria, which lack a methyl-directed nicking system.

278 just one strand of the recognition sequence, nicking the DNA rather than cleaving it.

279 -esterase activity, which is responsible for nicking the DNA strand to be transferred and for covalen

280 es that are typical of homing endonucleases, nicking the intron-minus polymerase genes in either host

281 c activity of APE1, which is responsible for nicking the phosphodiester backbone in DNA on the 5' sid

282 Nicking the target gene alone did not facilitate efficie

283 MutH initiates mismatch repair by nicking the transiently unmethylated daughter strand 5'

284 -dependent enhancer activation, based on DNA nicking to relieve torsional stress from eRNA synthesis.

285 d, topo I, and increased topo I-mediated DNA nicking under conditions of oxidative stress.

286 e either top-strand nicking or bottom-strand nicking variants from a randomly mutated SapI expression

287 sented the possibility that one, if not two, nicking variants might be created from SapI.

288 sites through mutagenesis, we have generated nicking variants of BtsCI that specifically nick the bot

289 icant amino acid sequence identity and their nicking variants were successfully isolated.

290 mutagenesis was carried out to isolate BsaI nicking variants with minimal double-stranded DNA cleava

291 d LAGLIDADG HEases, the engineering of their nicking variants, and the applications of NEases and nic

292 Top-strand specific nicking variants, Nt.BsrDI and Nt.BtsI, were successfull

293 e appropriate combinations of FokI and BtsCI nicking variants, we are able to generate long overhangs

294 The rate of nicking varies with the structure of the DNA but can occ

295 re repaired with high fidelity, simultaneous nicking via appropriately offset guide RNAs is required

296 ences are known to be unusually sensitive to nicking via the Fe2+-mediated Fenton reaction.

297 Arteriovenous nicking was also associated with prevalent (odds ratio =

298 The aberrant nicking was found to correlate with increased formation

299 Arteriovenous nicking was significantly associated with subcortical in

300 The effect of antigen nicking was then investigated by comparing the immunogen