马鹿β-防御素-1cDNA全长克隆、序列信息及表达分析

doi:10.11843/j.issn.0366-6964.2017.02.005

Abstract

Abstract:

This study aimed to study the gene structure and function of red deer β-defensin-1 (redBD-1) in order to better understand the gene's tissue expression pattern. The cDNA full-length sequence of redBD-1 gene was cloned from the red deer's lingual mucous membrane adopting the technology of PCR combining with rapid-amplification of cDNA ends (RACE), the sequence was analysed by bioinformatics, and the expression of the gene in different tissues was determined by Real-time quantitative PCR (RT-qPCR) technology. The results showed that the full-length sequence of redBD-1 gene was 455 bp, and the open reading frame (ORF) was 192 bp encoding 64 amino acids. The bioinformatics analysis indicated that theoretical molecular weight of redBD-1 protein was 6.94 ku containing 10 amino acid residues with positive charges without amino acid residue with negative charges, and its theoretical isoelectric point was 10.85. It could be predicted that redBD-1 protein had a secreting signal peptide structure, had no trans-membrane domain and mainly exereds the ectocellular physiological function; six conserved cysteine residues formed 3 intramolecular disulfide bonds with the connection of Cys1-Cys5, Cys2-Cys4 and Cys3-Cys6; the tertiary structure of maturation protein consisted of β-overlap, extension and random coils. Amino acid sequence of redBD-1 showed highest similarity with that of siBD-1 (98.4%), followed with BEBD (92.2%), HBD-2 (35.9%). RT-qPCR results indicated that redBD-1 was expressed in all detected organs, and the expression level was relatively higher in most organs of digestive system, respiratory system and reproductive system compared with a relatively lower expression level in parenchymatous organs like liver, kidney and spleen. This research could provide a theoretical basis for a better study of defensins gene functions and red deer mucosal immune system.

CLC Number:

S825
S813.1

TIAN Qiao-zhen, JIN Xin, ZHANG Man,CAI Shuo, LIU Jiao, WANG Yun-he, YANG Yin-feng. The Full-length Cloning, Sequence Information and Expression Analysis of Red Deer β-defensin-1 (redBD-1) cDNA[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 225-234.

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The Full-length Cloning, Sequence Information and Expression Analysis of Red Deer β-defensin-1 (redBD-1) cDNA

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