对一个包含ANT2及四个可能基因的135 kb Xq25酵母人工染色体进行有序鸟枪法测序，其中三个基因已通过EST匹配得到确认。
Ordered shotgun sequencing of a 135 kb Xq25 YAC containing ANT2 and four possible genes, including three confirmed by EST matches.
作者信息
Chen C N, Su Y, Baybayan P, Siruno A, Nagaraja R, Mazzarella R, Schlessinger D, Chen E
机构信息
Applied Biosystems Division, ACGT, Foster City, CA 94402, USA.
出版信息
Nucleic Acids Res. 1996 Oct 15;24(20):4034-41. doi: 10.1093/nar/24.20.4034.
Abstract
Ordered shotgun sequencing (OSS) has been successfully carried out with an Xq25 YAC substrate. yWXD703 DNA was subcloned into lambda phage and sequences of insert ends of the lambda subclones were used to generate a map to select a minimum tiling path of clones to be completely sequenced. The sequence of 135 038 nt contains the entire ANT2 cDNA as well as four other candidates suggested by computer-assisted analyses. One of the putative genes is homologous to a gene implicated in Graves' disease and it, ANT2 and two others are confirmed by EST matches. The results suggest that OSS can be applied to YACs in accord with earlier simulations and further indicate that the sequence of the YAC accurately reflects the sequence of uncloned human DNA.
摘要
已使用Xq25酵母人工染色体（YAC）底物成功进行了有序鸟枪法测序（OSS）。将yWXD703 DNA亚克隆到λ噬菌体中，并利用λ亚克隆的插入末端序列生成图谱，以选择要进行全测序的克隆的最小拼接路径。135038个核苷酸的序列包含整个ANT2 cDNA以及计算机辅助分析提示的其他四个候选序列。其中一个推定基因与格雷夫斯病相关基因同源，并且它、ANT2和另外两个基因通过EST匹配得到证实。结果表明，OSS可以按照早期模拟应用于YAC，并且进一步表明YAC的序列准确反映了未克隆的人类DNA的序列。
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