title: [Prokaryotic expression and identification of human papillomavirus type 16 E7 protein encoding gene].
abstract: OBJECTIVE
  To construct the prokaryotic expression plasmid pET32/E7 and express the human papillomavirus type 16 E7 protein in E. coli.
  
  
  METHODS
  HPV16 E7 gene was amplified by PCR. The amplified E7 fragment was inserted into the plasmid pET32a (+) that was digested with BamH I and Hind III. The recombinant plasmid pET32/E7 was transformed into E. coli JM109 which was selected with ampicillin. The positive clones containing recombinant plasmid pET32/E7 were verified by BamH I and Xho I digestion, and then sequenced. HPV16 E7-TRX recombinant protein expression in the E. coli BL21(ED3) was identified by SDS-PAGE and Western blot.
  
  
  RESULTS
  The prokaryotic recombinant plasmid pET32/E7 was successfully constructed. The BL21(DE3) transformed recombinant plasmid pET32/E7 had expressed HPV16 E7-TRX recombinant protein effectively. Under the conditions of 1 mmol/L IPTG and 30 degrees C, the amount of HPV16 E7-TRX recombinant protein was about 30% of bacterial total proteins.
  
  
  CONCLUSIONS
  The construction of the prokaryotic recombinant plasmid pET32/E7 and the successful expression of the recombinant protein HPV16 E7-TRX would strongly promote the research of the biological properties and the transformational mechanism of the HPV16 E7 protein on the specific cells.
authors.affiliation: Department of Microbiology, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
authors.forename: Qiao-Fa
authors.initials: QF
authors.lastname: Shi
authors.forename: Bao-Ning
authors.initials: BN
authors.lastname: Wang
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Wei
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Zhang
authors.forename: Kang
authors.initials: K
authors.lastname: Cao
authors.forename: Zhong-Hua
authors.initials: ZH
authors.lastname: Jiang
authors.forename: Ming-yuan
authors.initials: MY
authors.lastname: Li
mesh_terms: D000328:Adult; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D052162:Human papillomavirus 16 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D050725:Papillomavirus E7 Proteins / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011387:Prokaryotic Cells / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D050725:Papillomavirus E7 Proteins; D011994:Recombinant Proteins