[The targeting study of human serum albumin gene].
To generate transgenic porcine which expresses human serum albumin (HSA), the HSA gene targeting vector was constructed with HSA cDNA as the gene of interestand partial porcine serum albumin (PSA) gene as homologous arms which respectively were 7.2 kb 5' regulation sequence and 2.8 kb genomic sequence from the first intron to the fourth intron. The resistant gene neo was inserted into intron 1 and tk was ligated to the 3' end of the construct. Linearized targeting construct DNA was introduced into the fibroblast cells obtained from porcine fetus by electroporation. The positive-negative selection was performed and survival clones were screened by PCR and Southern blot. Three colonies with correct homologous recombination were obtained. Our results set a good basis for the establishment of transgenic porcine by gene target and nuclear transfer methods.
Sun
Shi-Hui
SH
Institute of Biotechnology, Academy of Military Medical Science, Beijing 100071, China. shsun916@sohu.com
Zhou
Yan-Rong
YR
Chen
Hong-Xing
HX
Lin
Yan-Li
YL
Huang
Pei-Tang
PT
Deng
Ji-Xian
JX
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002470:Cell Survival / Q000235:genetics; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018274:Electroporation; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D007621:Karyotyping; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012709:Serum Albumin / Q000235:genetics*; D013552:Swine; D014162:Transfection / Q000379:methods*
D012709:Serum Albumin