[Culture of HSV-2 and cloning of specific fragment of the gG-2 gene].
OBJECTIVE
To clone the glycoprotein G gene and its specific fragment with high conservation and antigenicity by culturing and amplifying herpes simplex virus type 2 and extracting its whole genome.


METHODS
We obtained a great deal of suspension with HSV-2 virus after infecting the cultured Hela cells with HSV-2 virus, extracted the whole genome of the virus by the phenol-chloroform method, and amplified the US4 gene coding gG-2 by PCR. Then we selected the specific target fragment according to the amino acid sequence alignment of the gG-2 gene and cloned it with the designed primers with restricted endonuclease sites.


RESULTS
We successfully obtained a lot of suspension with HSV-2 virus, and cloned the gG-2 gene from the whole genuine and its specific target fragment. Sequencing showed that both the sequences were identical with those printed in the GenBank.


CONCLUSIONS
It is feasible to obtain the virus genome and specific fragment of the gG-2 gene from virus-infected cells, especially for HSV-2 virus with relatively stable hereditary trait. It has prepared the ground for further constructing the expression plasmid of the specific fragment, expressing related proteins and identifying their antigenicity.
Gao
Jian
J
1. Institute of Life Science, Nanjing Normal University, Nanjing, Jiangsu 210046, China.
Huang
Yu-feng
YF
Cao
Jing
J
Wang
Hao-yang
HY
Lu
Jin-chun
JC
D000956:Antigens, Viral / Q000235:genetics*; D003001:Cloning, Molecular; D004279:DNA, Viral; D006367:HeLa Cells; D018258:Herpesvirus 2, Human / Q000235:genetics*; D006801:Humans; D014759:Viral Envelope Proteins / Q000235:genetics*; D014776:Virus Cultivation
D000956:Antigens, Viral; D004279:DNA, Viral; D014759:Viral Envelope Proteins