›› 2017, Vol. 29 ›› Issue (9): 1474-1481.DOI: 10.3969/j.issn.1004-1524.2017.09.08

• Animal Science • Previous Articles     Next Articles

Molecular cloning and expression analysis of FAM134B gene in Tibetan sheep

LI Wenli1, 2, DU Xiaohua2, 3, LIU Xia1, 2, *, GAO Jingbo1, 2, SHEN Chaochao1, 2, ZHANG Chao1, 2, LIU Wei1, 2   

  1. 1.College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
    2. Gansu Key Laboratory of Herbivorous Animal Biotechnology, Lanzhou 730070, China;
    3. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2016-12-26 Online:2017-09-20 Published:2017-09-27

Abstract: The gene of FAM134B(family with sequence similarity 134, member B) was cloned by RT-PCR. The bioinformatics analysis was carried out by using biological software. The expression level of FAM134B was also studied by real-time fluorescence quantitative PCR. The full length of FAM134B gene was 1 144 bp (GenBank accession number: KX580309), open reading frame 1 071 bp, encoding 356 amino acids. The phylogenetic tree showed that the Tibetan sheep FAM134B had a close relationship with the sheep and goat. The molecular formula of FAM134B protein in Tibetan sheep was C1717H2696N448O572S12, its molecular weight was 39151.72 u, the theory isoelectric point was 4.49, the extinction coefficient was 24 450. The instability index was 43, the aliphatic index was 80.31, and the grand average hydropathicity was -0.422. It was unstable and soluble acidic protein. The secondary structure of FAM134B was mainly composed of α-helices and random coil and extended chain, with α-helices was 45.51%, random coil was 41.85% and extension chains was 12.64%, which belongs to mixed type of protein. The results of subcellular localization and function showed that the frequencies of FAM134B protein in endoplasmic reticulum and cytoplasmic membrane were 30.4% and 21.7%. It mainly plays a role in protein transport and binding(77.3%). Phylogenetic tree showed that the amino acid sequence of FAM134B showed high homology with sheep and goat. qRT-PCR results showed that the expression of FAM134B gene was the highest in the stomach and lowest in the duodenum. The results can provide a theoretical basis for further study on the structure and physiological functions and genetic characteristics of FAM134B gene.

Key words: Tibetan sheep, FAM134B gene, CDS cloning, bioinformatics, expression

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