Construction and identification of lentiviral vector encoding human survivin gene

doi:10.3969/j.issn.2095-4344.2014.11.019

Abstract

Abstract:

BACKGROUND: Inhibiting the apoptosis of intervertebral disc cells can postpone the degenerative process of intervertebral disc. Survivin has a strong function of regulating cell proliferation and anti-apoptosis.

OBJECTIVE: To construct and identify the lentiviral vector encoding survivin gene of human.

METHODS: The survivin gene of human (BIRC5) was synthesized through the gene synthesis technology, amplified by PCR and analyzed by electrophoresis. The target gene was cloned into lentiviral expression plasmid to obtain the recombinant lentiviral vector Lenti-BIRC5. After transformation into competent E. coli cells, the candidate clones were identified by PCR firstly. The positive clones were identified by gene sequencing. The lentivirus plasmid containing target gene was transfected into 293T cells, and the expression of recombinant lentiviral vector Flag-Survivin fusion protein was detected through western blot analysis.

RESULTS AND CONCLUSION: The PCR results of electrophoresis and DNA sequencing showed that lentiviral vector containing human survivin gene was constructed successfully. Western blot analysis results showed that the target gene was transfected successfully and over-expressed in cultured cells. The lentiviral expression vector of human survivin gene Lenti-BIRC5 was constructed successfully, which lays a foundation for the study addressing the anti-apoptotic effects of survivin on human nucleus pulposus cells.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: lentivirus, osteocytes, cell apoptosis, intervertebral disk

CLC Number:

R318

Zhao Liang, Zhang Guo-qing, Ma Xue-xiao, Yang Kun, Hu You-gu, Chen Bo-hua. Construction and identification of lentiviral vector encoding human survivin gene[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(11): 1755-1760.

Figures/Tables 4

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