高通量测序技术结合正向遗传学手段在基因定位研究中的应用.pdfVIP

Hereditas (Beijing) 2015 年 8 月, 37(8): 765 ―776 综 述 高通量测序技术结合正向遗传学手段在基因定位研究 中的应用 陆才瑞，邹长松，宋国立 中国农业科学院棉花研究所，棉花生物学国家重点实验室，安阳 455000 摘要 : 传统的利用正向遗传学方法的基因定位一般是通过构建遗传连锁图谱进行的，该过程步骤繁琐、耗时耗 力，很多情形下定位精确度低、区间大。随着高通量测序技术的快速发展以及测序成本的不断降低，多种简单 快捷的利用测序手段定位基因的方法被开发出来，包括对突变体基因组直接测序定位、突变体材料构建混池测 序定位和遗传分离群体测序构建图谱定位等，还可以对转录组和部分基因组进行测序定位。这些方法可以在核 苷酸水平鉴定突变位点，并已推广到复杂的遗传背景中。近期报道的一些测序定位甚至是在不依赖于参考基因 组序列、遗传杂交和连锁信息的情况下完成的，这使得很多非模式物种也能开展正向遗传学研究。本文就这些 新技术及其在基因定位中的应用进行了综述。 关键词 : 高通量测序；基因定位；正向遗传学 Recent progress in gene mapping through high-throughput sequencing technology and forward genetic approaches Cairui Lu, Changsong Zou, Guoli Song State Key Laboratory of Cotton Biology, Institute of Cotton Research of Chinese Academy of Agricultural Sciences, Anyang 455000, China Abstract: Traditional gene mapping using forward genetic approaches is conducted primarily through construction of a genetic linkage map, the process of which is tedious and time-consuming, and often results in low accuracy of mapping and large mapping intervals. With the rapid development of high-throughput sequencing technology and decreasing cost of sequencing, a variety of simple and quick methods of gene mapping through sequencing have been developed, including direct sequencing of the mutant genome, sequencing of selective mutant DNA pooling, genetic map construction through sequencing of individuals in population, as well as sequencing of transcriptome and partial genome. These methods can be used to identify mutations at the nucleotide level and has been applied in complex genetic background. Recent reports have shown that sequencing mapping could be even done without the reference of genome sequence, hybridization, and genetic linkage information, which made it possible to perform forward genetic study in many non-model species. In this review, we summarized these new te