[The cloning and prokaryotic expression of human tumor necrosis factor like protein].
OBJECTIVE
To identify and clone the gene encoding human TNFLP (tumor necrosis factor like protein) for some functional study on TNFLP.


METHODS
The 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.


RESULTS
The 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.


CONCLUSIONS
Tissue expression spectrum of TNFLP and prokaryotic expression of TNFLP have been done, which establish the base for the functional analysis of TNFLP.
Hu
Xiao-yan
XY
National Laboyatory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005, China.
Zhou
Yan
Y
Dai
Yun
Y
Chen
Jian-meng
JM
Zhang
Bin
B
Peng
Xiao-zhong
XZ
Yuan
Jian-gang
JG
Qiang
Bo-qin
BQ
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005333:Fetus; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011387:Prokaryotic Cells / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D017421:Sequence Analysis; D017385:Sequence Homology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics*
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha