[Expression and purification of GST-HPV16E7 fusion protein].
OBJECTIVE
To construct a recombinant prokaryotic expression vector efficiently expressing HPV16E7 and purify GST-HPV16E7 fusion protein.


METHODS
The HPV16E7 gene obtained from a local cervical cancer patient was cloned into vector pGEX-4T-1, to generate the recombinant named as pGEX-4T-1-HPV16E7. The recombinant plasmid was transformed into E. coli BL21. After inducing with IPTG the HPV16E7 fusion protein was analyzed by SDS-PAGE and Western blot. B-PER GST Fusion Protein Purification Kit was appkied to purify GST-HPV16E7 fusion protein.


RESULTS
The recombinant plasmid pGEX-4T-1-HPV16E7 was successfully constructed. Highly expressed and purified GST-HPV16E7 fusion protein was obtained. The specificity of fusion protein was verified by SDS-PAGE and Western blot.


CONCLUSIONS
GST-HPV16E7 fusion protein was successfully expressed and purified.
Zhang
Jian
J
Department of Obstetrics and Gynecology, West China Second Hospital, Sichuan University, Chengdu 610041, China.
Wang
Tao
T
Xi
Ming-Rong
MR
Han
Min
M
Chen
Yue
Y
D001483:Base Sequence; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000235:genetics; D052162:Human papillomavirus 16 / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D018307:Neoplasms, Squamous Cell / Q000473:pathology / Q000821:virology; D050725:Papillomavirus E7 Proteins / Q000096:biosynthesis* / Q000235:genetics; D030361:Papillomavirus Infections / Q000821:virology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D002583:Uterine Cervical Neoplasms / Q000473:pathology / Q000821:virology
D050725:Papillomavirus E7 Proteins; D011993:Recombinant Fusion Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D005982:Glutathione Transferase