畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (9): 1678-1685.doi: 10.11843/j.issn.0366-6964.2015.09.026

• 研究简报 • 上一篇    下一篇

STARD3NL基因克隆和表达谱分析

邢晋祎1*,贾坤航1,李艳平2   

  1. (1.临沂大学生命科学学院,临沂 276005;2.山东农业大学动物科技学院,泰安 271018)
  • 收稿日期:2014-09-28 出版日期:2015-09-23 发布日期:2015-09-23
  • 通讯作者: 邢晋祎,E-mail:xingjinyi@lyu.edu.cn
  • 作者简介:邢晋祎(1968-),男,山东曹县人,博士,副教授,主要从事动物分子遗传学研究
  • 基金资助:

    山东省自然科学基金(ZR2013CL012);国家自然科学基金(31372333);临沂大学博士启动基金(BS08019)

Molecular Cloning and Expression Patterns Analysis of the Porcine STARD3 N-terminal like Protein(STARD3NL) Gene

XING Jin-yi1* ,JIA Kun-hang1,LI Yan-ping2   

  1. (1.College of Life Science,Linyi University,Linyi 276005,China;2.College of Animal Science and Veterinary Medicine,Shandong Agricultural University,Tai’an 271018,China)
  • Received:2014-09-28 Online:2015-09-23 Published:2015-09-23

摘要:

本研究旨在克隆猪STARD3NL (STARD3 N-terminal like protein) 基因,对其基因结构进行分析,并研究该基因在猪不同组织中的表达特性。采用5′和3′-RACE以及RT-PCR方法克隆猪STARD3NL基因,利用生物信息学方法预测蛋白结构,应用qRT-PCR技术检测组织表达特性。结果表明,猪STARD3NL基因的cDNA序列长度为1 319 bp(GenBank登录号:HQ634946),编码235个氨基酸,该氨基酸序列与其他物种相比一致性较高;进化树分析表明,猪STARD3NL氨基酸序列与牛和绵羊的关系较近,与斑马鱼的进化关系最远。STARD3NL氨基酸序列包含保守的MENTAL结构域,为疏水蛋白,包含4个跨膜螺旋区;二级结构主要是α-螺旋,占60%;该蛋白不存在信号肽,不是分泌蛋白;亚细胞定位显示,该蛋白存在于内质网的概率为39.1%。qRT-PCR分析表明,STARD3NL基因在所检测猪的14种组织均有表达,在肝中表达水平最高,在眼肌中的表达水平最低;莱芜猪和大长猪被圆环病毒感染后,STARD3NL基因在2种猪肺组织中的表达水平差异不显著(P>0.05)。研究结果将为进一步研究STARD3NL基因的结构和功能以及猪抗病育种提供资料。

Abstract:

The aim of this study was to clone porcine STARD3NAL(STARD3 N-terminal like protein),and analyze the structure of STARD3NAL,and investigate its expression in porcine different tissues.The porcine STARD3NAL gene was cloned by 5′,3′-RACE and RT-PCR,the protein structure of porcine STARD3NL was analyzed by bioinformatics methods.The expression patterns in different tissues were detected by real-time RT-PCR.The results showed that the porcine STARD3NL cDNA was 1 319 bp(GenBank accession No.:HQ634946) in length,which coded 235 amino acids and shared highly identity with those of other species.The phylogenetic tree analysis indicated the porcine STARD3NL was closely related to cattle and sheep,but distantly related to the zebrafish.The porcine STARD3NL protein contained conserved MENTAL domain and was hydrophobicity protein and there were 4 transmembrane helices in the STARD3NL protein.The secondary structure of STARD3NL protein showed that the STARD3NL protein fold for 60% into α-helix.This polypeptide contained no signal peptide in its amino acids,and was a nonsecretory protein.The residing probability of STARD3NL in the endoplasmic reticulum was 39.1%.By real-time PCR,mRNA expression of porcine STARD3NL were detected in all 14 tissues investigated,and STARD3NL mRNA expression levels in liver were the highest and in longissimus dorsi were the lowest.In addition,no significant difference of STARD3NL mRNA expression levels in lung tissues was detected between Laiwu pigs(LW) and YL pigs(Yorshire♂×Landrace♀) in infected circovirus group(P>0.05).These results will provide the data for further research of STARD3NL gene structures and function,breeding for disease resistance.

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