AIM： To clone human liver special F protein and to express it in a prokaryotic system. METHODS： Total RNA was isolated from human liver tissue and first-strand cDNA was reverse transcribed using the PCR reverse prim...AIM： To clone human liver special F protein and to express it in a prokaryotic system. METHODS： Total RNA was isolated from human liver tissue and first-strand cDNA was reverse transcribed using the PCR reverse primer. Following this, cDNA of the F protein was ligated into the clone vector pUCm-T. The segment of F protein＇s cDNA was subdoned into the expression vector pET-15b and transformed into E. coli BL21 （DE3） pLyss. Isopropy-β-D-thiogalactoside （IPTG） was then used to induce expression of the target protein. RESULTS： The cDNA clone of human liver special F protein （1134bp） was successfully produced, with the cDNA sequence being published in Gene-bank： DQ188836. We confirmed the expression of F protein by Western blot with a molecular weight of 43 kDa. The expressed protein accounted for 40% of the total protein extracted. CONCLUSION： F protein expresses cDNA clone in a prokaryotic system, which offers a relatively simple way of producing sufficient quantities of F protein and contributes to understanding the principal biological functions of this protein.展开更多
基金Supported by a grant from the Tianjin Science and Technology Committee, No. 023802911
文摘AIM： To clone human liver special F protein and to express it in a prokaryotic system. METHODS： Total RNA was isolated from human liver tissue and first-strand cDNA was reverse transcribed using the PCR reverse primer. Following this, cDNA of the F protein was ligated into the clone vector pUCm-T. The segment of F protein＇s cDNA was subdoned into the expression vector pET-15b and transformed into E. coli BL21 （DE3） pLyss. Isopropy-β-D-thiogalactoside （IPTG） was then used to induce expression of the target protein. RESULTS： The cDNA clone of human liver special F protein （1134bp） was successfully produced, with the cDNA sequence being published in Gene-bank： DQ188836. We confirmed the expression of F protein by Western blot with a molecular weight of 43 kDa. The expressed protein accounted for 40% of the total protein extracted. CONCLUSION： F protein expresses cDNA clone in a prokaryotic system, which offers a relatively simple way of producing sufficient quantities of F protein and contributes to understanding the principal biological functions of this protein.
