Construction and identification of a prokaryotic expression plasmid encoding HPV16E7-HSP70 fusion gene / 中国耳鼻咽喉头颈外科
Chinese Archives of Otolaryngology-Head and Neck Surgery
;
(12)2006.
Article
in Chinese
| WPRIM
| ID: wpr-527089
ABSTRACT
OBJECTIVE To construct a prokaryotic expression plasmid encoding HPV16E7-HSP70 fusion gene for further study on the immunity of HPV16E7- HSP70 fusion protein against laryngeal carcinoma. METHODS HPV16E7 was PCR-amplified,digested by NheI and SacI,and ligated into pET28a. HSP70 was cloned into pGEMTeasy,then recut from the vector by SalI and NotI and ligated into pET28a-HPV16E7. PCR amplification, restrict enzyme digestion, DNA sequencing, IPTG induction and Western Blot were used to identify the recombinant plasmid. RESULTS Double digestion and PCR amplification of the recombinant plas- mid have shown that the size of the inserted fragment is as expected. Sequence analysis has demonstrated that the inserted fragment encodes for the HPV16E7- HSP70 fusion gene. IPTG induction and Western Blot have shown that the fusion protein is expressed suc- cessfully in the prokaryotic expression plasmid. CONCLUSION The recombinant prokaryotic expression plasmid pET28a-HPV16E7-HSP70 has been con- structed successfully.
Full text:
Available
Index:
WPRIM (Western Pacific)
Type of study:
Diagnostic study
Language:
Chinese
Journal:
Chinese Archives of Otolaryngology-Head and Neck Surgery
Year:
2006
Type:
Article
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