Basic Research
Copyright ©2007 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. Mar 28, 2007; 13(12): 1799-1804
Published online Mar 28, 2007. doi: 10.3748/wjg.v13.i12.1799
Cloning and expression of special F protein from human liver
Shu-Ye Liu, Xin-Da Yu, Chun-Juan Song, Wei Lu, Jian-Dong Zhang, Xin-Rong Shi, Ying Duan, Ju Zhang
Shu-Ye Liu, Wei Lu, Jian-Dong Zhang, Xin-Rong Shi, Ying Duan, Tianjin Third Central Hospital, Tianjin Artificial Cell Laboratory, Tianjin 300170, China
Shu-Ye Liu, Xin-Da Yu, Chun-Juan Song, Ju Zhang, Nankai University, Biology Science Institute, Tianjin 300071, China
Author contributions: All authors contributed equally to the work.
Supported by a grant from the Tianjin Science and Technology Committee, No. 023802911
Correspondence to: Shu-Ye Liu, Tianjin Third Central Hospital, Tianjin 300170, China. lshye@163.com
Telephone: +86-22-24384350
Received: November 13, 2006
Revised: November 17, 2006
Accepted: February 8, 2007
Published online: March 28, 2007
Abstract

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 subcloned 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.

Keywords: F protein; Liver; Gene-cloning; Protein expression; Prokaryotic system