染色体替换系的方案构建及评价

doi:10.3969/j.issn.1674-5817.2015.06.010

实验动物与比较医学 ›› 2015, Vol. 35 ›› Issue (6): 478-483.DOI: 10.3969/j.issn.1674-5817.2015.06.010

染色体替换系的方案构建及评价

赵莹^1,2, 赵丽亚^1,2, 晁天柱³, 张蓉^1,2, 邢正弘^1,2, 陈国强^1,2, 肖君华³

1. 上海西普尔-必凯实验动物有限公司, 上海 201203;
2. 上海实验动物研究中心, 上海 201203;
3. 东华大学生物科学与技术研究所, 上海 201620

收稿日期:2015-06-05 出版日期:2015-12-25 发布日期:2015-12-25
作者简介:赵莹(1982-), 女, 助研, 硕士, 从事群体遗传学及药物毒性安全评价研究。E-mail: 12414863@qq.com。
基金资助:
上海市科学技术委员会科研计划项目(12140900400、13140900302)

Construction and Evaluation of Chromosome Substitution Strains

ZHAO Ying^1,2, ZHAO Li-ya^1,2, CHAO Tian-zhu³, ZHANG Rong^1,2, XING Zheng-hong^1,2, CHEN Guo-qiang^1,2, XIAO Jun-hua³

1. Sino-British SIPPR/BK Laboratory Animal LTD., CO, Shanghai 201203, China;
2. Shanghai Laboratory Animal Research Center, Shanghai 201203, China;
3. Institute of Biological Sciences and Biotechnology, Donghua University, Shanghai 201620, China

Received:2015-06-05 Online:2015-12-25 Published:2015-12-25

摘要/Abstract

摘要： 目的为缩短复杂性状相关基因精细定位时间,构建染色体替换系(CSSs),并对构建方案进行评价。方法以供体小鼠作为母本、受体C57BL/6小鼠作为父本进行杂交, 获得N1代小鼠。将N1代雄性小鼠与C57BL/6雌性小鼠杂交, 获得N2代小鼠。选择N2代1号染色体非重组的雄性小鼠与C57BL/6雌性小鼠回交, 获得N3代回交小鼠; 通过短串联重复序列(STR)与单核苷酸多态性(SNP)对每代一定数量的子代进行遗传分析, 依次回交10代, 选择N10代1号染色体未重组的样本, 雌雄交配, 挑选1号染色体为纯合且来自供体品系的子代小鼠, 用于繁殖纯合后代。通过基因芯片扫描鉴定替换系的纯合度,并与期望值比较。结果根据构建的CSSs方案, 得到2个不同的CSSs, 通过基因芯片扫描得到的替换系纯合度与期望值接近。结论构建的CSSs方案可完成(部分)近交系小鼠品系目标染色体替换, 供体品系背景基因组中99.59%以上已被受体C57BL/6小鼠替换,证明构建的CSSs方案可行。

关键词: 染色体替换系(CSSs), 方案构建, 评价, 非重组率

Abstract: Objectives To shorten the fine-structure mapping time, construct and evaluate the programme of chromosome substitution strains (CSSs) . Methods The rst step required making hybrids between C57BL/6 male mice and donor female mice. N1 hybrids male mice were backcrossed to host C57BL/6 female mice. Progeny with a non-recombinant chromosome derived from donor strain, in this case chromosome 1, were identified in this and subsequent backcrosses. N2 Male mice were backcrossed to host C57BL/6 female mice at each generation. A certain number of progeny per generation was screened with short tandem repeat (STR) and single nucleotide polymorphism (SNP). These mice were backcrossed to host C57BL/6 at each generation until tenth generation. At the tenth backcross generation males and females with the non-recombinant chromosome derived from donor mice were intercrossed. Progeny of this intercross that were homozygous (homosomic) for chromosome 1 were used to propagate the homosomic strain. The homozygosity of CSSs was identified through scanning the whole genomic by microarray to compare with expected proportion. Results We have constructed two CSSs by the programme that had been constructed. The actual homozygosity of progeny with microarray was approaching the expected. Conclusion The constructed CSSs confirmed the replacement of target chromosome and over 99.59% chromosome segments from host C57BL/6. This breeding program to generate a CSSs panel proved to be feasible.

Key words: Chromosome substitution strains (CSSs), Construct the programme, Discuss, Rate of non-recombination

中图分类号:

Q95-33

赵莹, 赵丽亚, 晁天柱, 张蓉, 邢正弘, 陈国强, 肖君华. 染色体替换系的方案构建及评价[J]. 实验动物与比较医学, 2015, 35(6): 478-483.

ZHAO Ying, ZHAO Li-ya, CHAO Tian-zhu, ZHANG Rong, XING Zheng-hong, CHEN Guo-qiang, XIAO Jun-hua. Construction and Evaluation of Chromosome Substitution Strains[J]. Laboratory Animal and Comparative Medicine, 2015, 35(6): 478-483.

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染色体替换系的方案构建及评价

Construction and Evaluation of Chromosome Substitution Strains

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