| Autor: |
De-liang, Li, Wen-li, Ma, Yong-xia, Shi, Ling, Li, Bao, Zhang, Wen-ling, Zheng |
| Rok vydání: |
2007 |
| Předmět: |
|
| Zdroj: |
Nan fang yi ke da xue xue bao = Journal of Southern Medical University. 26(12) |
| ISSN: |
1673-4254 |
| Popis: |
To construct an eukaryotic expression plasmid containing gp120 gene of HIV-1 subtype B and obtain gp120 gene expression in HepG2 cells.According to the published gp120 gene sequence in Genbank, a pair of primers was designed and synthesized. The PCR amplification product of gp120 gene was cloned into pMD-18T vector using TA cloning followed by BamHI and XhoI digestion and sequence analysis. The target gene was then subcloned into a highly efficient eukaryotic expression vector pcDNA3.1 (+). The recombinant plasmid was sequenced and identified by restrictive endonuclease digestion, and transfected into HepG2 cells via liposome. The expression of gp120 gene was analyzed by RT-PCR and Western blotting, respectively.Restriction endonuclease digestion and sequence analysis verified successful construction of the recombinant vector pcDNA3.1(+)/gp120. The target fragment gp120 was identical with U26942 in Genbank, and the expression of gp120 gene was detected in the lysate of the transfected HepG2 cells by RT-PCR and Western blotting.The eukaryotic expression plasmid for gp120 has been constructed successfully, which is capable of stable expression in HepG2 cells. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
|
