›› 2015, Vol. 42 ›› Issue (4): 830-837.doi: 10.16431/j.cnki.1671-7236.2015.04.009

Previous Articles     Next Articles

Cloning and Sequence Analysis of Buffalo BDNF Gene and Investigation of its Expression Pattern in Different Tissues

DU Feng-jiao, LIU Xiao-lin, WU Zhu-lian, LI Xiao-kai, QU Chun-feng, ZHAO Xin, LEI Chuan, RUAN Qiu-yan, LU Feng-hua, SHI De-shun   

  1. State Key Laboratory of Conservation and Utilization of Subtropical Agro-bioresources, Guangxi University, Nanning 530004, China
  • Received:2014-08-11 Online:2015-04-20 Published:2015-05-05

Abstract: In this study, the CDS sequence of buffalo BDNF gene was cloned, and then the homology, evolutive tree of BDNF amino acid sequence and the physicochemical characteristics, secondary and tertiary structures of BDNF protein were analyzed and predicted by bioinformatics techniques, also, the expression pattern of BDNF mRNA in different tissues of fetal and adult buffalo was investigated by QRT-PCR. The results showed that with RT-PCR a 800 bp buffalo BDNF gene fragment was cloned and sequenced, including the whole CDS of 753 bp (coded 250 amino acid). The multiple sequence alignment result showed that, the BDNF gene in buffalo shared 99%, 94%, 93%, 90%, 90% and 89% similar nucleotide sequence with that of Bos taurus, Sus scrofa, Canis lupus familiaris, Homo sapiens, Equus caballus and Mus musculus, respectively. The evolutive tree analysis showed that BDNF gene was highly conservative in evolution. The molecular weight and isoelectric point of BDNF protein were predicted as 28 173.36 u and 9.12, respectively. The secondary structure of BDNF protein was composed of several α-helix, β-sheet, T-turn and randon coil structures, and the tertiary structure of BDNF protein was composed of several α-helix and 3 pairs of antiparallel β-sheet structures and so on, which configuring an active center aslo called the NGF functional domain. The QRT-PCR result showed that BDNF mRNA completely expressed in heart, lung, kidney, brain, muscle, ovary and testis in both of fetal and adult buffalo, but the mRNA expression in adult buffalo was higher than that of fetal buffalo.

Key words: buffalo; BDNF; cloning and analysis; expression pattern

CLC Number: