Study On The Structure And Expression In Gene Of Beijing Fatty Chicken And Wuzhishan Mini-Swine

In this paper,extraction the total RNA of Beijing Fatty Chicken and Wuzhishan Mini-Swine's liver as a template,clone target genes to PGEM-T Easy through RT-PCR,construct Beijing Fatty Chicken's cloning vector of PGEM-T-IL-15,PGEM-T-IL-18,PGEM-T-CRABP,PGEM-T-LPL,PGEM-T-TBP,PGEM-T-musclin,PGEM-T-L-FABP and Wuzhishan Mini-Swine's cloning vector of PGEM-T-L-FABP,PGEM-T-SLA-DQA.construct Beijing Fatty Chicken and Wuzhishan Mini-Swine's expression vector pEGFP-N3-L-FABP through the method of double restriction enzyme,and expression in target cells.The results are below:1.This study clone the gene of IL-15,IL-18,CRABP,LPL,TBP,musclin and L-FABP from Beijing Fatty Chicken and the gene of L-FABP,SLA-DQA from Wuzhishan Mini-Swine, submit gene sequence to GenBank and obtain embody's number.2.This study successfully established the target cell lines through direct adherent culture metheods,The cells sample numbers of 44,conserced numbers are 210,each one density is about 3～5×106cell/ml. after recovery the cells are typical fibroblast,according to the requirement of cell lines to biological analysis,The cell lines accord with the request of ATCC. It provides well target cells for gene expression.3.This study construct Beijing Fatty Chicken's expression vector pEGFP-N3-L-FABP through XhoI and EcoRI method of double restriction enzyme and expression in target cells.construct Wuzhishan Mini-Swine's expression vector pEGFP-N3-L-FABP through BamhI and XhoI method of double restriction enzyme and expression in target cells.It establish a good foundation for somatic cell nuclear transfer.