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发明公开
CN103409408A 快速获取鹅铁蛋白重链基因编码区序列及定量检测其表达的方法及其引物 无效 - 撤回

  • 专利标题： 快速获取鹅铁蛋白重链基因编码区序列及定量检测其表达的方法及其引物
  • 专利标题（英）： Method for rapidly obtaining coding region sequence of goose ferritin heavy-chain gene and quantitatively detecting expression of gene, and primers thereof
  • 申请号： CN201310303514.X
    申请日： 2013-07-19
  • 公开(公告)号： CN103409408A
    公开(公告)日： 2013-11-27
  • 发明人: 康波 , 姜冬梅 , 白林 , 王迅 , 赵玲 , 王林杰 , 王艳 , 何珲 , 马容
  • 申请人： 四川农业大学
  • 申请人地址： 四川省雅安市雨城区新康路46号
  • 专利权人： 四川农业大学
  • 当前专利权人： 四川农业大学
  • 当前专利权人地址： 四川省雅安市雨城区新康路46号
  • 主分类号： C12N15/10
  • IPC分类号： C12N15/10 ;  C12Q1/68 ;  C12N15/11
快速获取鹅铁蛋白重链基因编码区序列及定量检测其表达的方法及其引物
摘要：
本发明公开了一种快速获取铁蛋白重链基因编码区序列及定量检测其表达的方法及其引物，包括以下步骤：提取鹅组织总RNA并反转录；设计FTH编码区序列引物FTH-L（SEQ ID NO:1-2）和检测FTH表达规律的引物FTH-S（SEQ ID NO:3-4）；通过RT-PCR扩增，产物回收纯化，目的基因连接和转化，阳性菌落的筛选和测序，获取FTH编码区序列，测序鉴定荧光定量PCR引物。该技术成本低、效率高，只需一次克隆即可获得完整编码区序列；并可快速、准确检测鹅不同组织和不同发育时期样品中FTH的表达规律，为阐明鹅FTH结构和功能，以及鹅机体内铁代谢机制的研究奠定了理论基础。
摘要（英）：

The invention discloses a method for rapidly obtaining the coding region sequence of a goose ferritin heavy-chain gene and quantitatively detecting the expression of the gene, and primers thereof. The method comprises the following steps: extracting the total RNA from goose tissues, carrying out inverse transcription, and designing an FTH coding region sequence primer FTH-L (SEQ ID NO:1-2) and an FTH expression rule detection primer FTH-S (SEQ ID NO:3-4); and carrying out RT-PCR amplification, recovering and purifying the obtained product, carrying out target gene connection and conversion, and screening and sequencing a positive bacterial colony to obtain the FTH coding region sequence and the sequencing identification fluorescence quantitative PCR primers. The method has the advantages of low cost, high efficiency, obtaining of the complete coding region sequence only through one-time cloning, and rapid and accurate detection of the expression rule of the FTH in samples of different tissues and different growth periods, and lays a theoretic foundation for the clarification of the FTH structure and function, and the researches of the iron metabolism mechanism in the goose.

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