Cloning, expression and protein structure prediction of a novel SCF-TPO fusion gene
- VernacularTitle:重组人干细胞因子／血小板生成素融合基因的克隆、表达及其蛋白质结构预测
- Author:
Nan LIU
;
Yongzhi XI
;
Siqi GUO
;
Yuying SUN
;
Zhihong YUAN
;
Jianwu CUI
;
Caixia XI
;
Fei LIANG
;
Fanhua KONG
- Publication Type:Journal Article
- Keywords:
Stem cell factor;
Thrombopoitin fusion protein;
Prokaryotic expression;
Protein structure
- From:
Chinese Journal of Pathophysiology
2000;0(10):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To obtain the high expression of recombinant human stem cell factor - thrombopoitin (SCF-TPO) fusion gene and predict its structure property. METHODS: Tour primers were designed according to known sequence of TPO and SCF to amplify the functional amino acid domain of TPO and SCF by RT- PCR, respectively from fetus hepatocytes. The expression plasmid pET32a/SCF- TPO was constructed by VOE gene fusion technique and expressed in BL21(DE3)plysS. The fusion protein property, such as second structure, flexibility, and hydrophilicity were predicted by DS Gene and Protscale software. RESULTS: The expression vector, pET32a/SCF - TPO was constructed and the high expression of the SCF/TPO fusion protein was obtained, with the expression amount of up to 40% of the total cellular protein. DS Genel .5 and Protscale predict no new antigenicity in fusion protein, and the second structure and ioelectric point have no changes except four amino acids change in first structure. There are high flexibility and low hydrophilieity in the linker peptide. CONCLUSION: High expression of SCF- TPO fusion protein has been obtained and protein prediction shows that the fusion protein design is reasonable, which lay foundation for further study of biological fundation of SCF - TPO fusion protein.
