[Molecular cloning of human IL-7cDNA and construction of eukaryotic vector expressing hIL-7]
OBJECTIVE: To construct a vector expressing eukaryotic human interluken-7(hIL-7). METHODS: hIL-7 DNA was identified and cloned (cDNA) from human spleen tissue using reverse transcription polymerase chain reaction (RT-PCR). We incorporated the cDNA into the pMD18-T plasmid. The pMD18-T plasmid was then inserted into a dual expression vector (prokaryotic and eukaryotic) pBK-CMV and called pBK-CMV-hIL-7. We used pBK-CMV-hIL-7 vector to infect E.coli DH5alpha. The expression of the recombinant hIL-7 protein (rhIL-7) by E.coli DH5alpha was analyzed using SDS-PAGE and western blot testing. RESULTS: The genetically engineered E.coli DH5alpha did express rhIL-7 confirmed by western blot. CONCLUSION: The successful construction of genetically engineered eukaryotic gene for hIL-7 was done, This will enable further research into therapeutic uses for hIL-7.
Cheng
Bei
B
The Affiliated Obstetrics and Gynecology Hospital, College of Medical Sciences, Zhejiang University, Hangzhou 310006, China.
Yie
Da-Feng
DF
Xie
Xing
X
Chen
Huai-Zeng
HZ
Lu
Wei-Guo
WG