The cloning and prokaryotic expression of human tumor necrosis factor like protein.
- Author:
Xiao-yan HU
1
;
Yan ZHOU
;
Yun DAI
;
Jian-meng CHEN
;
Bin ZHANG
;
Xiao-zhong PENG
;
Jian-gang YUAN
;
Bo-qin QIANG
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Animals; Base Sequence; Brain; metabolism; Cloning, Molecular; DNA, Complementary; genetics; Escherichia coli; genetics; Fetus; Humans; Mice; Molecular Sequence Data; Prokaryotic Cells; metabolism; Recombinant Fusion Proteins; biosynthesis; genetics; Sequence Analysis; Sequence Homology; Tumor Necrosis Factor-alpha; biosynthesis; genetics
- From: Acta Academiae Medicinae Sinicae 2002;24(3):246-249
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify and clone the gene encoding human TNFLP (tumor necrosis factor like protein) for some functional study on TNFLP.
METHODSThe full-length cDNA of TNFLP was isolated from fetal brain cDNA library. Several kinds of software were used to analyze nucleotide sequence and amino acid sequence of TNFLP. TNFLP mRNA distribution was identified by Northern blot. TNFLP-C and TNFLP-N were expressed in E. coli with GST expression system.
RESULTSThe cDNA of human TNFLP was 2,112 bp, which encoded protein of 208 amino acid. Hydrophobility analysis found there were two hydrophobility regions of human TNFLP. TNFLP-C (112-207 amino acid) and mouse TNF-alpha were homologous. The identity of their amino acid sequence was 42%. Moreover, both of them had a motif-TYKRL. TNFLP was located in chromosome 16. Human TNFLP was widely expressed in various human tissues. Northern blot showed TNFLP was highly expressed in heart, brain and spleen, only one transcript can be seen. GST-TNFLP-C and GST-TNFLP-N fusion proteins were obtained.
CONCLUSIONSTissue expression spectrum of TNFLP and prokaryotic expression of TNFLP have been done, which establish the base for the functional analysis of TNFLP.
