B6-Co小鼠Ankrd55和Ddx4突变候选基因克隆及测序

doi:10.3969/j.issn.1674-5817.2012.05.001

实验动物与比较医学 ›› 2012, Vol. 32 ›› Issue (5): 371-375.DOI: 10.3969/j.issn.1674-5817.2012.05.001

• 华东地区第12 届实验动物科学学术交流会优秀论文选刊 • 下一篇

B6-Co小鼠Ankrd55和Ddx4突变候选基因克隆及测序

李瑶¹, 吴刘成¹, 卢泽艳¹, 郑良凤¹, 葛良玉¹, 邵义祥^1,2

1.南通大学实验动物中心, 南通 226001;
2.南通大学比较医学研究所, 南通 226001

收稿日期:2012-01-31 出版日期:2012-10-25 发布日期:2012-10-25
作者简介:李瑶(1988-),女,硕士研究生,专业方向:疾病动物模型。E-mail:ly3322332@163.com
基金资助:
国家自然科学基金项目(30671081);江苏省自然科学基金项目(BK2010279);南通大学自然科学基金项目(11Z052)

Mutation Screening of Two Candidate Genes in B6-Co Mouse Affected with Cornea Opacity: Ankrd55 and Ddx4

LI Yao¹, WU Liu-cheng¹, LU Ze-yan¹, ZHENG Liang-feng¹, GE Liang-yu¹, SHAO Yi-xiang^1,2

1. Laboratory Animals Centre, Nantong University, Nantong 226001, China;
2. Comparative Medicine Institute, Nantong University, Nantong 226001, China

Received:2012-01-31 Online:2012-10-25 Published:2012-10-25

摘要/Abstract

摘要： 目的对B6-Co小鼠Ankrd55和Ddx4基因进行克隆测序分析,寻找是否存在突变位点。方法以小鼠Ankrd55和Ddx4基因的mRNA序列设计引物,以mRNA为模板,采用RT-PCR和PCR技术分段进行目的基因扩增,将目的片段连接在T载体上,转化至感受态细胞,筛选阳性克隆,提取质粒DNA分子,电泳检测,EcoRΙ酶切释放目的片段,测序、分析、比对。结果 Ddx4基因位于小鼠13号染色体第113412349 的碱基由C转换成A,导致编码氨基酸的密码子改变,产物脯氨酸(P)变成谷氨酰胺(Q)。结论 Ddx4基因发生单碱基突变,表明该突变可能与B6-Co小鼠的EOB表型相关,但是有待于进一步研究。

关键词: B6-Co小鼠, 角膜混浊, PCR体外扩增, 基因克隆

Abstract: Objective To find mutational site of Ankrd55 and Ddx4 gene, clone and sequence these gene in B6-Co mutant strain mouse with cornea opacity phenotype. Methods The primers were designed according to mRNA. The target gene was amplified by PCR and RT-PCR in which the PCR templates were mRNA, and then recombinant vectors were transformed to competent cells after DNA fragments were connected to the T vector. The positive clones were selected and plasmid DNA extraction was detected by electrophoresis. The fragments by EcoRI digestion for identification of positive clones and at the end sequencing. Results The C base in sequence of Ddx4 chromosome 13 113 412 349 was replaced by A by sequence alignment with genome data base, with coding protein Pro replacement by Gln. Conclusion There is a single base mutation in Ddx4 gene, and the mutation may relate to the EOB phenotype of B6-Co mice. And it needs to be studied further.

Key words: B6 - Co mice, Corneal turbidity, PCR amplification, Gene clone

中图分类号:

Q785Q95-33

李瑶, 吴刘成, 卢泽艳, 郑良凤, 葛良玉, 邵义祥. B6-Co小鼠Ankrd55和Ddx4突变候选基因克隆及测序[J]. 实验动物与比较医学, 2012, 32(5): 371-375.

LI Yao, WU Liu-cheng, LU Ze-yan, ZHENG Liang-feng, GE Liang-yu, SHAO Yi-xiang. Mutation Screening of Two Candidate Genes in B6-Co Mouse Affected with Cornea Opacity: Ankrd55 and Ddx4[J]. Laboratory Animal and Comparative Medicine, 2012, 32(5): 371-375.

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B6-Co小鼠Ankrd55和Ddx4突变候选基因克隆及测序

Mutation Screening of Two Candidate Genes in B6-Co Mouse Affected with Cornea Opacity: Ankrd55 and Ddx4

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