Construction of recombinant retroviral vector with rat Islet-1 gene
- VernacularTitle:Islet-1基因逆转录病毒表达载体的构建
- Author:
Jiamei LIU
;
Dong CHEN
;
Xiaoting MENG
- Publication Type:Journal Article
- Keywords:
insulin gene enhancer binding protein 1;
clone;
recombinant retroviral expression vector
- From:
Journal of Jilin University(Medicine Edition)
2006;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To isolate the rat insulin gene enhancer binding protein 1(Islet-1)gene and construct plEGFP-C1-Islet-1 recombinant retroviral expression vector.Methods The cDNA encoding the rat Islet-1 gene was isolated by RT-PCR method,the cDNA was first cloned into PGET-1 TA vector to facilitate the sequence and then subcloned into the retroviral vector plEGFP-C1.plEGFP-C1-Islet-1 was transfected into PA317 packaging cells with lipofectamine 2000.Transformants were selected in medium containing G418.Results A 1 050 bp DNA fragment was obtained by RT-PCR;plEGFP-C1-Islet-1 recombinant retroviral expression vector was identified by restrictive enzymes digestion,PA317 cells transfected with recombinant vector expressed enhancer green fluorescent protein(EGFP).Conclusion The gene encoding the rat Islet-1 is obtained and plEGFP-C1-Islet-1 expression vector is constructed successfully.
