Cloning and expression of special F protein from human liver

• Published in: World journal of gastroenterology : WJG Vol. 13; no. 12; pp. 1799 - 1804
• Main Authors: Liu, Shu-Ye, Yu, Xin-Da, Song, Chun-Juan, Lu, Wei, Zhang, Jian-Dong, Shi, Xin-Rong, Duan, Ying, Zhang, Ju
• Format: Journal Article
• Language: English
• Published: United States Tianjin Third Central Hospital, Tianjin Artificial Cell Laboratory, Tianjin 300170, China 28.03.2007; Nankai University, Biology Science Institute, Tianjin 300071, China%Nankai University, Biology Science Institute, Tianjin 300071, China%Tianjin Third Central Hospital, Tianjin Artificial Cell Laboratory, Tianjin 300170, China; Baishideng Publishing Group Co., Limited
• Subjects: Basic Research; cDNA; Cloning, Molecular - methods; DNA, Complementary - genetics; Escherichia coli; F蛋白; Gene Expression Regulation; Genetic Vectors; Humans; Liver - cytology; Liver - metabolism; Proteins - genetics; Proteins - metabolism; RNA, Messenger - genetics; 人类肝脏; 基因克隆; 基因表达; F protein; Protein expression; Prokaryotic system; Gene-cloning; Liver
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v13.i12.1799

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.