Cloning and expression of special F protein from human liver.
OBJECTIVE
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-beta-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.


CONCLUSIONS
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.
Liu
Shu-Ye
SY
Tianjin Third Central Hospital, Tianjin Artificial Cell Laboratory, Tianjin 300170, China. lshye@163.com
Yu
Xin-Da
XD
Song
Chun-Juan
CJ
Lu
Wei
W
Zhang
Jian-Dong
JD
Shi
Xin-Rong
XR
Duan
Ying
Y
Zhang
Ju
J
D003001:Cloning, Molecular / Q000379:methods*; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006801:Humans; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics
D018076:DNA, Complementary; C521996:F protein, human; D011506:Proteins; D012333:RNA, Messenger