[Chicken SREBP1 gene antiserums preparation and its tissue expression characterization]. 2010

Li Wang, and Wei Na, and Yu-Xiang Wang, and Yan-Bo Wang, and Ning Wang, and Yu-Mao Li, and Hui Li
College of Animal Science and Technology, Northeast Agricultural University, Harbin 150030, China. wangli930@yahoo.com

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.

UI MeSH Term Description Entries
D007106 Immune Sera Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen. Antisera,Immune Serums,Sera, Immune,Serums, Immune
D009928 Organ Specificity Characteristic restricted to a particular organ of the body, such as a cell type, metabolic response or expression of a particular protein or antigen. Tissue Specificity,Organ Specificities,Specificities, Organ,Specificities, Tissue,Specificity, Organ,Specificity, Tissue,Tissue Specificities
D002645 Chickens Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA. Gallus gallus,Gallus domesticus,Gallus gallus domesticus,Chicken
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D004926 Escherichia coli A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc. Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D005786 Gene Expression Regulation Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation. Gene Action Regulation,Regulation of Gene Expression,Expression Regulation, Gene,Regulation, Gene Action,Regulation, Gene Expression
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D000918 Antibody Specificity The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site. Antibody Specificities,Specificities, Antibody,Specificity, Antibody
D000939 Epitopes Sites on an antigen that interact with specific antibodies. Antigenic Determinant,Antigenic Determinants,Antigenic Specificity,Epitope,Determinant, Antigenic,Determinants, Antigenic,Specificity, Antigenic

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