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人类11号染色体的序列标签位点图谱。

A sequence-tagged site map of human chromosome 11.

作者信息

Smith M W, Clark S P, Hutchinson J S, Wei Y H, Churukian A C, Daniels L B, Diggle K L, Gen M W, Romo A J, Lin Y

机构信息

Molecular Genetics Laboratory, Salk Institute for Biological Studies, La Jolla, California 92037.

出版信息

Genomics. 1993 Sep;17(3):699-725. doi: 10.1006/geno.1993.1392.

Abstract

We report the construction of 370 sequence-tagged sites (STSs) that are detectable by PCR amplification under sets of standardized conditions and that have been regionally mapped to human chromosome 11. DNA sequences were determined by sequencing directly from cosmid templates using primers complementary to T3 and T7 promoters present in the cloning vector. Oligonucleotide PCR primers were predicted by computer and tested using a battery of genomic DNAs. Cosmids were regionally localized on chromosome 11 by using fluorescence in situ hybridization or by analyzing a somatic cell hybrid panel. Additional STSs corresponding to known genes and markers on chromosome 11 were also produced under the same series of standardized conditions. The resulting STSs provide uniform coverage of chromosome 11 with an average spacing of 340 kb. The DNA sequence determined for use in STS production corresponds to about 0.1% (116 kb) of chromosome 11 and has been analyzed for the presence of repetitive sequences, similarities to known genes and motifs, and possible exons. Computer analysis of this sequence has identified and therefore mapped at least eight new genes on chromosome 11.

摘要

我们报告了370个序列标签位点(STS)的构建情况,这些位点在标准化条件下可通过PCR扩增检测到,并且已在区域上定位到人类11号染色体。通过使用与克隆载体中存在的T3和T7启动子互补的引物直接从黏粒模板进行测序来确定DNA序列。通过计算机预测寡核苷酸PCR引物,并使用一系列基因组DNA进行测试。通过荧光原位杂交或分析体细胞杂种板,将黏粒区域定位在11号染色体上。在相同的一系列标准化条件下,还产生了与11号染色体上已知基因和标记相对应的其他STS。所得的STS提供了对11号染色体的均匀覆盖,平均间距为340 kb。用于生成STS的DNA序列对应于11号染色体的约0.1%(116 kb),并且已分析了重复序列的存在、与已知基因和基序的相似性以及可能的外显子。对该序列的计算机分析已在11号染色体上鉴定并因此定位了至少八个新基因。

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