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E . coli cryptic miniplasmid p 15A 에서 유래하는 플라스미드 pACYC 184 의 ssi 시그날에 관한 연구
박정동,Sakai Hiroshi,Tohru Komano ( Jeong Dong Bahk,Hiroshi Sakai,Tohru Komano ) 생화학분자생물학회 1990 BMB Reports Vol.23 No.1
Single-strand DNA initiation (ssi) signals are DNA requirements for initiation of DNA synthesis on the single-strand DNA templates. For primary screening of ssi signal function, we used plaque morphology method. Plasmid pACYCl84 derived from E. coli minicryptic plasmid p15A had only one ssi signal which consists of 119-nt stretch. This segment played critical roles in ss DNA phage growth activity and performed primase-dependent replication instead of RNA polymerase-dependent one. Three kinds of energetically stable stem and loop structures were expected. Especially, the orientation and location of this 119-nt stretch, when compared with those of the origin of p15A, favor that this ssi signal might be taken part in the lagging strand replication of plasmid pACYCl84.
E . coli cryptic miniplasmid p 15A 에서 유래하는 플라스미드 pACYC 184 의 ssi 시그날에 관한 연구
박정동,Sakai, Hiroshi,Komano, Tohru 생화학분자생물학회 1993 BMB Reports Vol.17 No.3
Single-strand DNA initiation (ssi) signals are DNA requirements for initiation of DNA synthesis on the single-strand DNA templates. For primary screening of ssi signal function, we used plaque morphology method. Plasmid pACYCl84 derived from E. coli minicryptic plasmid p15A had only one ssi signal which consists of 119-nt stretch. This segment played critical roles in ss DNA phage growth activity and performed primase-dependent replication instead of RNA polymerase-dependent one. Three kinds of energetically stable stem and loop structures were expected. Especially, the orientation and location of this 119-nt stretch, when compared with those of the origin of p15A, favor that this ssi signal might be taken part in the lagging strand replication of plasmid pACYCl84.
CHUNG,SOO-YEOL,YOSHISUE,HAJIME,SAKAI,HIROSHI,KAWAMUKAI,MAKOTO,CHOI,YONG-LARK,KOMANO,TOHRU,CHO,MOO-JE 東亞大學校附設遺傳工學硏究所 1996 遺傳工學硏究 Vol.- No.3
Cyclic AMP receptor protein (CRP) binds to cAMP and regulates the expression of many genes negatively or positively in Escherichia coli (1,6,9,10). Mutants carrtying crp^*, in which CRP^* can actively affect gene expression without binding cAMP, were isolated by many groups (2,7,8). Most crp^* mutations were isolated as lactose-fermentative genes differently. For example, crp^*, which was isolated by Aiba et al., induced expression of the lac genes, but not of the mal genes in the absence of cAMP or cGMP (2). We have cloned at least 12 different E. coli genes. The cloning strategy has been described(11). Chromosomal DNA fragments of E. coli were inserted into pKK223-4, a multicopy expression vector, followed by introduction into the E. coli crp^* cya strain MK2001, and maltose-fermentative clones were selected. The genes were designated sfs1 throught sfs12(sugar fermentaion stimulation). Previously, sfs7(5) and sfs1(11) were analyzed. Althought crp^* is probably involved in some way in the transcriptional regulation of sfs genes, nothing has been mentioned for sfs1 and sfs7 from this point of view in these previous studies, In this study, another gene(sfs4) located at 53min on the E. coli chromosome is analyzed. We have found that maltose was fermented in the absence of cAMP in a crp^*1 cya strain when the sfs4 gene was overexpressed under the tac promoter and even when the gene was expressed under the intrinsic promoter. We have analyzed the transcription of sfs4.