Preliminary Study on Transformation of Human canstatin Gene to the Novel Bioreactor——Dunaliella salina
- VernacularTitle:人canstatin基因转化新型生物反应器——杜氏盐藻(Dunaliella salina)的初步研究
- Author:
Shu-Ying FENG
;
Hui-Hui GU
;
Hong-Tao LIU
;
Le-Xun XUE
;
- Publication Type:Journal Article
- Keywords:
Human canstatin Transformation Novel bioreactor Dunaliella salina
- From:
China Biotechnology
2006;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
The human canstatin cDNA was amplified by RT-PCR and then directionally cloned into pU? expression vector. The recombinant pU?-Can vector was connected with the screening marker (bar box), to construct a eukaryotic expression vector called pU?-Can-Bar. This expression vector was introduced into the D.salina by glass beads method. The screening culture of transformants of D.salina was performed in solid media containing 5 ?g/ml PPT, and the analyses of transformants were carried out through PCR and Southern blot. PCR results revealed that specific 700 bp products were detected in the different transformants of D.salina but not in negative control. Southern blot analysis further demonstrated that human canstatin gene was integrated into the D.salina genome. Moreover, the results of genetic stability analyses of transformants demonstrated that canstatin gene was stably inherited in the D.salina transformants. The successful preparation of the D.salina transformants will provide the experimentation evidence for producing canstatin protein cosmically by using the D.salina bioreactor and give a better prophase work basis for clinic application of canstatin protein early.
