Goose LB-FABP Gene Cdna Cloning And Effect Of Over-Feeding On Expression Patterns Of LB-FABP, APO-b And APO-A1in Goose Liver

FABP, Apo-A1and Apo-B genes have been proven to be important for lipid metabolism and played key roles in lipid anabolism, fat deposition, cholesterol transporting and energy homeostasis regulation. In this study we chose10Landes geese as experiment materials, which were divided into over-feeding group and control group. Total RNA was extracted from liver, followed by the synthesis of cDNA first strand. The CDS(coding sequence) sequence of FABP in goose liver was cloned with RACE and RT-PCR. The bioinformatics analysis of the sequence was done. The information about the construction and properties of this protein were predicted by other software. These would faciliate further study for function and expression of FABP in goose liver. At meantime, in order to probe into the influence of over-feeding treatment on gene expression of FABP, ApoB and ApoAl in Landes goose, SYBR Green Ⅰ real-time quantitative PCR was developed to analyze the expression characterization of FABP, Apo-B and Apo-A1genes. Some study found that the ratio of Apo-B/Apo-A1in plasma is a better index assessing atherosclerosis, diabete Ⅱ and insulin-resistence than either of them used alone. The ratio of Apo-B/Apo-A1in mRNA level is the ratio of the ratio of Apo-B to reference gene to that of Apo-A1. The results are as follows:1. cDNA sequences of goose Lb-FABP cloning and analysisThe FABP cDNA sequence cloned by RACE was532bp in length, which was submitted to GenBank(Accession No. KF039686). Using bioinformatics network resources and relevant software, we predicted that the protein sequence is126aa in length, after a20aa signal peptide predicted was deduced. It was predicted to be a basic hydrophilic protein, with high affinity with lipid, which can exist stablely in body. It was located in cytoplasm by analyzing for subcellular localization, including several potential phosphorylation sites. It belongs to cytosolic fatty-acid binding (FABP family). The secondary structure and3D structure prediction suggested that this protein has a typical fold of the FABP family in which10strands of antiparallel β-sheet surround the hydrophobic ligand binding cavity and two short a-helices are located between the firstand second strands. Comparison of the putative amino acid of porcine orthologs with that of other species showed that goose liver FABP shared62%,69%,82%,94%,94%and88%homology with Lb-FABP of Danio rerio (AAI64928.1), Xenopus (Silurana) tropicalis (XP002938785.1), Taeniopygia guttata (XP002196447.1), Gallus gallus (NP989965.1), Anas platyrhynchos (AEH03008.1) and Epinephelus coioides (AEU04719.1) respectively. We can classify FABP in goose liver as Lb-FABP, which was coded by Fabp10. Lb-FABP may be the only FABP in goose liver;2. Expression change after overfeeding of Lb-FABP, Apo-A1and Apo-BQuantitative real-time analysis reveals that after over-feeding treatment, mRNA level of goose Lb-FABP decreased(P>0.05). The regulation mechanism was unknow, The decrease of Lb-FABP can decrease absorption of fatty acid into hepatocyte, which in turn protected liver. mRNA level of goose liver Apo-B decreased(P>0.05), while mRNA level of goose liver Apo-A1increased (P>0.05). The ratio was significantly decreased (P<0.05), indicating that after overfeeding, elimination of cholesterol adhering to vascular wall and cholesterol transportation decrease, protecting cardiovascular from lesion. These gene expression changes showed a resistence to overfeeding and formation of Fatty liver. Apo-B/Apo-A1is a promising breeding index to create more healthy animal breeds for its excellence at assessing cardiovascular health.