| 摘要: |
| 采用RT-PCR和快速扩增cDNA末端(rapid amplification of cDNA ends, RACE)技术首次克隆了翘嘴鲌 TMEM-57蛋白基因的cDNA全序列, 该序列全长为2822bp, 其中5′-UTR区93bp, 3′-UTR区731bp, 开放阅读框(open reading frame, ORF)为1998bp, 编码665个氨基酸。NJ法系统进化分析将同科或属中TMEM-57基因分成一个分支, 表现出种属间的高度保守性。利用荧光定量PCR(qPCR)技术检测了不同组织的表达量, 发现在卵巢中表达量最高, 其次为心脏和脑。用直接测序法对TMEM-57基因部分区段单核苷酸多态性(SNP)进行了检测, 寻找到11个新的SNP位点。 |
| 关键词: 翘嘴鲌, TMEM-57, 荧光定量PCR, 单核苷酸多态性 |
| DOI：10.11693/hyhz201206024024 |
| 分类号: |
| 基金项目:国家科技支撑计划, 2012BAD25B06 号 |
附件 |
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| CLONING, TISSUE EXPRESSION AND SINGLE NUCLEOTIDE POLYMORPHISM ANALYSIS OF CULTER ALBURNUS TMEM-57 |
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QI Peng-Zh1, GUO Bao-Ying2, XIE Cong-Xin1, WU Chang-Wen2, LU Shi-Min1, DUAN You-Jian1
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1.College of Fisheries, Huazhong Agricultural University;2.National Research Centre of Mariculture Equipments and Engineering, Technology, Marine Science College of Zhejiang Ocean University
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| Abstract: |
| In this paper, a 2822bp full-length cDNA sequence of TMEM-57 gene from Culter alburnus was obtained with RT-PCR and rapid amplification of cDNA ends (RACE) technique. It consists of a 93bp 5′untranslated region (UTR), an 1998bp open reading frame (ORF) and a 731bp 3’UTR. The translated protein is composed of 665 amino acids. Neighbor-Joining (NJ) tree suggested that the TMEM-57 gene is highly conservative between species clustering the same species. After detected the gene expression quantity in tissues with real-time fluorescence quantitative PCR (qPCR), expression quantity in the ovary is considered highest, then the heart and brain. Certain fragments of TMEM-57 are inspected for single nucleotide polymorphism (SNP) by sequencing of PCR production, eleven novel SNP loci are detected. |
| Key words: Culter alburnus, TMEM-57, Real-time fluorescence quantitative PCR, Single nucleotide polymorphism |
