ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2012, Vol. 43 ›› Issue (5): 684-692.doi:

• 遗传繁育 • Previous Articles     Next Articles

Gene Clone, Subcellular Localization of Expression Products of HFABPand the Preparation of Transgenic Mice in Xuhuai Goat

YIN Yanhui1，WEI Guanghui1，LI Wei1，ZHU Caiye1，ZHANG Yani1，
DU Lixin2，CAO Wenguang2，LI Bichun1*   

  1. 1. Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science & Technology, Yangzhou University, Yangzhou 225009, China; 2. Institute of Animal Science，Chinese Academy of Agricultural Sciences，Beijing 100193,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2012-05-28 Published:2012-05-28
  • Contact: LI Bichun

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Abstract: The purpose of this study was to clone hearttype fatty acid binding protein (HFABP) gene cDNA of Xuhuai goat, and to explore its bioinformatics function and the possibility of preparation of transgenic animals among heterogeneous species. The subcelluar location HFABP was detected by EGFP fusion protein and its expression was observed in vitro. Reverse transcription PCR (RTPCR) technology was used to clone the HFABP gene cDNA of Xuhuai goat, its biological information characteristics was analyzed by online software, then the expression vector pEGFPHFABP was constructed. The transfection of goat fibroblasts (GEF) was performed by Liposomes (LTX), and fluorescence was observed under inverted microscope after 48 h. The RTPCR was conducted to detect mRNA expression of HFABP in GEF. The pEGFPHFABP was injected into mouse testicular and its expression was detected at the level of DNA and protein. The complete CDS size of HFABP was 402 bp, encoding 133 amino acids with GenBank accession number (AY466498.1). The HFABP cDNA coding sequence was compared with the corresponding regions of human, chicken, brown rat, cow, wild boar, donkey and zebra fish, the similarity was 89%, 76%, 85%, 84%, 93%, 91%, 70%, respectively, amino acid sequence homology was 90%, 79%, 88%, 97%, 95%, 94%, 72%, respectively. The signal peptide was not found in HFABP protein. The RTPCR results showed the HFABP mRNA expressed successfully in vitro. pEGFPHFABP was successfully constructed, and HFABP mRNA was expressed. The HFABP protein was localized in the cytoplasm which was in line with the result of online prediction. The gene can aslo be expressed in mice transiently and persistencely after intravenous and testicular injection. The HFABP gene cDNA of Xuhuai goat was cloned successfully, and it was conservative during the evolutionary process, there was no signal peptide in protein. The HFABP protein was located in the cytoplasm, and also could be expressed in mice successfully.