[Chicken SREBP1 gene antiserums preparation and its tissue expression characterization].
OBJECTIVE
The aim of this study was to prepare the antiserums against chicken sterol regulatory element binding protein1 (SREBP1), and to analyze the expression of SREBP1 in chicken tissues.


METHODS
The nuclear import sequence of SREBP1 was analyzed using the DNAStar programs to predict its major antigen epitopes, the fragment coding for SREBP1 major antigen epitopes (832-1 302 bp) was amplified by RT-PCR and inserted into pGEX-4T-1 to construct the expression vector pGEX-4T/SREBP1. The recombinant GST/SREBP1 was expressed in E.coli with IPTG induction and purified by Glutathione Sepharose 4B affinity chromatography. The purified recombinant GST/SREBP1 was used as immunogen in rabbits, and the titer and specificity of SREBP1 antiserums were detected by ELISA and Western blot. The tissue expression of chicken SREBP1 was analyzed with the antiserums.


RESULTS
The titer of the antiserum determined by ELISA was 1:102 400, and the antiserums exhibited a high specificity through Western blot. Tissue expression analysis showed that the expression of chicken SREBP1 was much higher in abdominal adipose tissue and heart, but lower in liver, muscle stomach, duodenum, spleen, kidney, and no signal was detected in breast and leg muscle. The SREBP1 expression characteristics between fat and lean broiler lines was also detected, and the result showed that SREBP1 expressed significantly higher in the abdominal adipose tissue of fat line than that in lean line (P<0.05).


CONCLUSIONS
The antiserum against chicken SREBP1 prepared in this study had high titer and specificity. SREBP1 was highly expressed in abdominal adipose tissue of broilers. Compared to lean broiler line, SREBP1 was expressed higher in the fat line.
College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China. wangli930@yahoo.com
Li
L
Wang
Wei
W
Na
Yu-Xiang
YX
Wang
Yan-Bo
YB
Wang
Ning
N
Wang
Yu-Mao
YM
Li
Hui
H
Li
D000818:Animals; D000918:Antibody Specificity; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics; D007106:Immune Sera / Q000276:immunology*; D009928:Organ Specificity; D051780:Sterol Regulatory Element Binding Protein 1 / Q000235:genetics* / Q000276:immunology*
D000939:Epitopes; D007106:Immune Sera; D051780:Sterol Regulatory Element Binding Protein 1