[Function of TTG1A in hepatic stellate cells].
OBJECTIVE
To investigate the biological functions of TTG1A in liver fibrosis.


METHODS
Yeast two-hybrid system was used to screen proteins associated with TTG1A. Briefly, the coding sequence of TTG1A was cloned into pGBKT7 vector, and the recombinant plasmid was transformed into yeast cells AH109 ( a type), then these cells were mated with yeast cells Y187 (a type) transformed with human leukocyte cDNA library plasmid pACT2. The obtained diploid yeast cells were plated on synthetic dropout nutrient medium containing X-alpha-gal for double selection. The plasmids from positive colonies were transformed into E.coli and sequenced.


RESULTS
The recombinant yeast expression vector pGBKT7-TTG1A was successfully constructed. Nineteen TTG1A binding proteins, including Homo sapiens major histocompatibility complex, class II DP beta 1 (HLA-DPb1), Homo sapiens ribosomal protein L30 (RPL30), Homo sapiens nucleophosmin Homo sapiens nucleobindin 2 (NUCB2), Homo sapiens ash2, variant Gaucher disease and variant metachromatic leukodystrophy, MORF4L1, Homo sapiens ubiquitin-conjugating enzyme E2L3 (UBE2L3), APOA1, Homo sapiens lectin, and galectin 1, were identified.


CONCLUSIONS
This study may help to elucidate the molecular function of TTG1A.
Xiao
Lin
L
Institute of Infectious Diseases, Beijing Ditan Hospital, Beijing 100015, China.
Cheng
Jun
J
Zhang
Li-ying
LY
Guo
Jiang
J
Hong
Yuan
Y
Zhang
Li-juan
LJ
Lun
Yong-zhi
YZ
Lan
Xian-yong
XY
Wu
Hui-juan
HJ
Zhang
Yue-xin
YX
Zhang
Jian-long
JL
Li
Yan
Y
D002352:Carrier Proteins / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015723:Gene Library; D005809:Genes, Regulator; D005822:Genetic Vectors; D055166:Hepatic Stellate Cells; D006801:Humans; D008103:Liver Cirrhosis / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids / Q000235:genetics; D012269:Ribosomal Proteins / Q000235:genetics; D015533:Transcriptional Activation; D053773:Transforming Growth Factor beta1 / Q000235:genetics; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D002352:Carrier Proteins; D018076:DNA, Complementary; D012269:Ribosomal Proteins; D053773:Transforming Growth Factor beta1