凡纳滨对虾BTF3基因的克隆及表达分析

摘要

通过电子克隆所得序列设计引物,采用RT-PCR方法首次克隆得到长655 bp的凡纳滨对虾通用转录因子3(Basic Transcription Factor3,BTF3)基因序列,其中包括597 bp的完整开放阅读框,共编码198个氨基酸残基,推算分子量约45.79 kDa,理论等电点为4.93.氨基酸序列分析表明,BTF3基因编码的氨基酸序列中具有保守的NAC结构功能域,利用实时荧光定量PCR检测凡纳滨对虾不同组织的表达情况发现,该基因在凡纳滨对虾组织中广泛表达,其中肌肉组织表达量最高,在肠中表达量最低.%A pair of primers were designed according to the sequence from electronic cloning, and a 6S5 bp cDNA of BTF3 in Utopenaeus vannamd was first obtained by RT-PCR. The cDNA contained a 597 bp open reading frame ( ORF) encoding a 198 amino acids pepfide. The molecular weight of BTF3 was 45.79 kDa, and its calculated isoelectric point was 4.93. Sequence alignment revealed that the amino acid sequence of BTF3 from different species were highly conserved and contained a NAC domain. Real-time PCR results showed that BTF3 mRNA could be detected in most of Utopenaeus vannamei tissues, and the highest expression level was observed in muscle, the lowest in intestine.