›› 2018, Vol. 45 ›› Issue (9): 2358-2367.doi: 10.16431/j.cnki.1671-7236.2018.09.002

Previous Articles     Next Articles

Cloning, Bioinformatics and Tissue Expression Analysis of SCP2 Gene in Dairy Buffalo

LIANG Shasha, PANG Chunying, DENG Tingxian, MA Xiaoya, LU Xingrong, DUAN Anqin, LIANG Xianwei   

  1. Guangxi Key Laboratory of Buffalo Genetics, Breeding and Reproduction, Guangxi Buffalo Research Institute, Chinese Academy of Agricultural Sciences, Nanning 530001, China
  • Received:2018-03-09 Online:2018-09-20 Published:2018-09-26

Abstract:

The study was aimed to clone sterol carrier protein 2 (SCP2) of buffalo,analyze the sequences by bioinformatics software,and measure the expression of SCP2 gene in different tissues of buffalo.The complete CDS of buffalo SCP2 gene was successfully cloned which was 1 632 bp in length and encoded 543 amino acids using the SCP2 gene (GenBank accession No.:NM_001033990.3) of Bos taurus as the seed sequence.The CDS of buffalo SCP2 gene showed 95.9%,93.4%,92.4%,89.4%,88.3%,86.3% and 86.9% identity with that of Bos taurus,Ovis aries,Capra hircus,Delphinapterus leucas,Homo sapiens,Felis catus familiaris and Canis lupus,and the results of phylogenetic tree showed that the nearest relationship existed between buffalo and Bos taurus.The results of amino acid sequence analysis showed that the formula of protein encoded by SCP2 gene in buffalo was C2602H4131N709O774S298, and the molecular weight,theory isoelectric point,instability index and average of hydropathicity was 58.66 ku,8.59,27.94 and -0.215 respectively,showing it was an alkaline,hydrophilic and stable protein.The secondary structure of SCP2 protein mainly consisted of α-helices (35.54%),random coil (48.99%) and extended strand (15.47%),which was consistent with the results of tertiary structure prediction.The results of subcellular localization showed that SCP2 protein was located in the cytoplasm (43.5%),peroxisome (21.7%),mitochondria (17.4%),nuclear (13.0%),and cytoskeleton (4.4%).There was no transmembrane domain and signal peptide,indicating it was an non secretory protein.There were 13 Ser,3 Thr and 2 Tyr which might be a protein kinase phosphorylation site.Prediction of protein binding sites showed that buffalo SCP2 protein contained 12 protein binding sites and 1 polynucleotide binding site.The Real-time PCR results showed that the expression of buffalo SCP2 gene in the liver was obviously higher than other tissues,which followed by mammary gland,lymph,kidney,large intestine,stomach,lung,spleen,ovary,pituitary,brain and heart ranged from high to low.This study provided the basis for further studies on the function of SCP2 gene.

Key words: buffalo; sterol carrier protein 2(SCP2); gene cloning; bioinformatics analysis; tissue expression

CLC Number: