Isolation, physical mapping and polymorphism of the porcine ferredoxin reductase (FDXR ) gene.
Isolation, physical mapping and polymorphism of the porcine ferredoxin reductase (FDXR ) gene.
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DOI：
10.1046/j.1365-2052.2002.00896_12.x
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发表时间：
2002-10
期刊：
影响因子：
2.4
通讯作者：
M. Yu;S. Zhao;M. Yerle;B. Liu;T. Xiong;K. Li
中科院分区：
文献类型：
--
作者：
M. Yu;S. Zhao;M. Yerle;B. Liu;T. Xiong;K. Li
来源/描述：利用Baskin和Pomp2引物对Roslin pigE BAC文库1进行聚合酶链反应（PCR）筛选，分离到一个含有猪胶原酶基因（CLPS）的BAC克隆（140E21）。通过序列分析证实了该基因在BAC克隆（140E21）中存在。对来自BAC克隆140E21的DNA扩增的PCR产物进行测序，发现与先前公布的序列相同（GenBank登录号：AF012819）。用HaeIII酶切BAC克隆（140E21）的DNA，亚克隆到pUC18 （Amersham Pharmacia Biotech, Little Chalfont, Bucks， UK）中。筛选转化子的CA / TG重复序列和潜在阳性序列。一个这样的亚克隆，虽然不包含CA重复，但发现包含一个polyA区域，随后是一个中断（GAAA） 5 （GenBank: AJ306320）。这个匿名的DNA序列与CLPS基因有物理上的联系，位点符号为S0628。引物设计在复杂重复区两侧，聚合酶链反应产物的多态性通过变性聚丙烯酰胺凝胶电泳证实。
Source⁄ description: A BAC clone (140E21) containing the porcine colipase gene (CLPS) was isolated after screening the Roslin pigE BAC library1 by polymerase chain reaction (PCR) with primers described by Baskin and Pomp2. The presence of the colipase gene in the BAC clone (140E21) was confirmed by sequence analysis. A PCR product amplified from DNA from BAC clone 140E21 was sequenced and found to be identical to the previously published sequence (GenBank accession number: AF012819). DNA from the BAC clone (140E21) was digested with HaeIII and subcloned into pUC18 (Amersham Pharmacia Biotech, Little Chalfont, Bucks, UK). Transformants were screened for CA⁄ TG repeats and potential positives sequenced. One such subclone, while not containing a CA repeat, was found to contain a polyA region followed by an interrupted (GAAA) 5 (GenBank: AJ306320). This anonymous DNA sequence, which is physically linked to the CLPS gene, has been assigned the locus symbol S0628. Primers were designed flanking the complex repeat region and the polymorphic nature of the PCR products demonstrated by denaturing polyacrylamide gel electrophoresis.