[Construction of eukaryotic expression vector for human angiopoietin 1 and its expression in the marrow mesenchymal stem cells of rabbit].
OBJECTIVE
To investigate the possibility of constructing eukaryotic expression vector for human angiopoietin 1 (hAng-1), transfecting it to bone marrow mesenchymal stem cells (MSCs) so as to repair bone defect.


METHODS
The eukaryotic expression vector pcDNA3-hAng-1 was constructed by recombinant DNA technique, transfected into MSCs by liposome DOTAP, and selected with G418. The hAng-1 expression of mRNA and protein was detected by reverse transcript-PCR and Western Blot.


RESULTS
After the recombinant eukaryotic expression vector for hAng-1 was digested with Xho-I and BamH-I, electrophoresis revealed 1.4 kb fragment for hAng-1 gene and 5.4 kb fragment for pcDNA3 vector. In the transfected MSCs, the mRNA and protein expression of hAng-1 gene were detected with reverse transcript-PCR and Western Blot.


CONCLUSIONS
The constructed eukaryotic expression vector hAng-1 could be expressed in the transfected MSCs, thus to provide the basis for bone repair with tissue engineering.
Zhang
Ye
Y
Department of Orthopaedic Surgery, Shanghai Sixth People's Hospital, Jiaotong University, Shanghai, 200233, PR China. zhangyeyun@yahoo.com.cn
Zeng
Bingfang
B
Zhang
Changqing
C
D042683:Angiopoietin-1 / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism*; D002478:Cells, Cultured; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011817:Rabbits; D014162:Transfection
D042683:Angiopoietin-1; D012333:RNA, Messenger