摘要
【目的】克隆延边黄牛转铁蛋白受体2(transferrin receptor 2,TFR2)基因,并分析该基因在不同组织器官中的表达分布。【方法】通过提取肝脏组织总RNA,采用RT-PCR及RACE方法克隆延边黄牛TfR2基因,采用生物信息学软件进行序列分析,用半定量PCR(SqRT-PCR)方法分析TfR2基因在不同组织器官中的表达分布规律。【结果】①成功克隆出了延边黄牛TfR2基因完整ORF区及3′UTR区(GenBank登录号:GU553087),该基因全长2 901 bp,ORF区2 412 bp,编码803个氨基酸;②延边黄牛TfR2基因核苷酸序列与其它物种的同源性在80%以上,不同物种间TfR2基因氨基酸序列,尤其是影响其功能的关键氨基酸位点高度保守;③各物种TfR2蛋白功能结构域同样高度保守;④牛TfR2基因主要在肝脏中表达,其它组织,如脾脏、心脏、肾脏和肠道(十二指肠)中也存在少量表达。【结论】不同物种间TfR2基因具有高度同源性,功能结构域及组织分布规律具有高度保守性。
【Objective】 The transferrin receptor 2（TfR2） gene was cloned from Yanbian Yellow cattle,the sequence was analyzed by bioinformatics software and the tissue expression distribution was tested by semi-quantitative reverse transcription and polymerase chain reaction（SqRT-PCR）.【Method】 The total RNA was extracted from Yanbian Yellow cattle liver.The TfR2 gene was cloned by reverse transcription polymerase chain reaction（RT-PCR） and rapid amplification of cDNA ends（RACE）.The TfR2 gene was analyzed by bioinformatic softwares and the TfR2 gene tissue distribution was tested by SqRT-PCR in the end.【Result】 The complete CDs and 3′ UTR region of TfR2 gene was cloned（GenBank accession： GU553087）.The TfR2 gene was 2901 bp in length and contained a 2412-nucleotides-long open reading frame（ORF） which encoded a protein of 803 amino acid residues.The sequence analysis result showed that Yanbian Yellow cattle TfR2 gene has over 80% homology with mammals in nucleotides,the amino acid sequences of TfR2 gene from different mammals were conserved,especially some amino acid sites which affect its function in human were highly conserved.The protein functional domain analysis showed that TfR2 gene from different mammals were highly conserved.The expression profiles analysis showed that the TfR2 gene was highly expressed in liver,constitutively expressed in spleen,heart,kidney and intestine（duodenum）.【conclusion】 The TfR2 gene from Yanbian Yellow cattle is highly homologous with other mammals,the functional domain and tissue distribution are highly conserved in different mammals.
出处
《中国农业科学》
CAS
CSCD
北大核心
2012年第10期2022-2030,共9页
Scientia Agricultura Sinica
基金
国家“863”计划项目(2008AA101010)
国家“十一五”科技支撑项目(2008BADB2B01)
