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合成寡核苷酸作为杂交探针的应用:人β2-微球蛋白克隆cDNA序列的分离

Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta 2-microglobulin.

作者信息

Suggs S V, Wallace R B, Hirose T, Kawashima E H, Itakura K

出版信息

Proc Natl Acad Sci U S A. 1981 Nov;78(11):6613-7. doi: 10.1073/pnas.78.11.6613.

Abstract

We have synthesized two sets of 15-base-long oligodeoxyribonucleotides corresponding to all possible coding sequences for a small portion of human beta 2-microglobulin. Labeled oligonucleotides were used as hybridization probes to screen bacterial clones containing cDNA sequences primed with oligo(dT) and inserted into the plasmid vector pBR322. One beta 2-microglobulin cDNA clone was detected in the 535 bacterial plasmid clones that were screened. The clone has been characterized by blotting and nucleotide sequence analysis. The cloned beta 2-microglobulin sequence contains 217 base pairs of the 3' untranslated region of the mRNA and 328 base pairs (97%) of the coding region.

摘要

我们合成了两组15个碱基长的寡聚脱氧核糖核苷酸,它们对应于人β2-微球蛋白一小部分的所有可能编码序列。标记的寡核苷酸用作杂交探针,以筛选含有由寡聚(dT)引发并插入质粒载体pBR322的cDNA序列的细菌克隆。在筛选的535个细菌质粒克隆中检测到一个β2-微球蛋白cDNA克隆。该克隆已通过印迹和核苷酸序列分析进行了表征。克隆的β2-微球蛋白序列包含mRNA 3'非翻译区的217个碱基对和编码区的328个碱基对(97%)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9432/349099/fad945ddbcc5/pnas00662-0076-a.jpg

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