Constructing lentivirus vector carrying human KiSS-1 gene cloning
- VernacularTitle:人KiSS-1基因克隆及其慢病毒载体的构建
- Author:
Bo YU
;
Li ZHANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(42):-
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Construction of lentivirus vector has the advantages of simplicity,it is regard as the most effective and successful method in transgene therapy. OBJECTIVE: To clone the metastasis suppressor gene KiSS-1 from human normal placenta tissue and construct its lentivirus vector. DESIGN,TIME AND SETTING: The open experiment was performed at the Laboratory of Fujian Normal University from September 2006 to December 2007. MATERIAL: The pNL-IRES2-EGFP vector was conservated by the Laboratory of Fujian Normal University. METHODS: Total RNA was extracted from human placenta tissue. The opening reading frame cDNA of KiSS-1 was isolated by using RT-PCR,and cloned into its lentiviral vector pNL-IRES2-EGFP to construct expression plasmid pNL-IRES2-EGFP-KiSS-1. MAIN OUTCOME MEASURES: Clone objective gene fragment of KiSS-1,restriction enzyme digestion and gene sequencing of the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 were observed in the study. RESULTS: The nucleotide sequence isolated from the recombinant plasmid pNL-IRES2-EGFP-KiSS-1 was confirmed the same as expected by restriction enzyme digestion and gene sequencing. CONCLUSION: The recombinant plasmid pNL-IRES2-EGFP-KiSS-1 has been constructed successfully.
