[Construction of high efficient eukaryotic expression recombinant on human hepatocyte DNA synthetic stimulated factor].
OBJECTIVE
To construct the eukaryotic expression recombining vector on human The pGEM-hepatocyte DNA synthetic stimulated factor (hHDSSF) with gene cloning.


METHODS
hHDSSF, a mid-recombining vector, was constructed by T-A cloning. After restriction endonuclease Not I digestion, the target fragment was subcloned into eukaryotic vector pcDNA3. 1hisB to construct the eukaryotic expression recombinant pcDNA3. 1hisB-HDSSF.


RESULTS
The forward insert recombinant pcDNA3. 1hisB-HDSSF was screened and obtained with restriction endonuclease Kpn I digestion and it was detected by DNA sequence analysis.


CONCLUSIONS
The eukaryotic expression recombinant pcDNA3. 1hisB-hHDSSF on hHDSSF is constructed successfully, which lays a foundation for building a stable eukaryotic expression cell strain and expressing hHDSSF.
Tang
Shi-gang
SG
Department of Infectious Diseases, Third Xiangya Hospital, Central South University, Changsha 410013, China.
Su
Xian-shi
XS
D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D017228:Hepatocyte Growth Factor / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D011994:Recombinant Proteins; C485002:hepatocyte DNA synthetic stimulated factor, human; D017228:Hepatocyte Growth Factor