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携带酵母核糖体DNA的大肠杆菌质粒内的缺失

Deletions within E. coli plasmids carrying yeast rDNA.

作者信息

Cohen A, Ram D

出版信息

Gene. 1978 Apr;3(2):135-47. doi: 10.1016/0378-1119(78)90057-4.

Abstract

Deletions occur in recombinant DNA plasmids that contain yeast ribosomal DNA (rDNA) inserted into the E. coli plasmids pSC101 and pMB9. Deletions within a pMB9 plasmid containing an insert longer than one tandem rDNA repeat apparently are due to homologous recombination because (1) all of the independently derived deletion products of this plasmid lost one complete rDNA repeat (8.6 kb) and retained only a single copy of the segment repeated at the ends of the original insert and (2) deletions were detected only when the insert had terminal redundancy. Deletions also occur within a pSC101 plasmid containing a tandem duplication of a segment (4.7 kb) including both pSC101 DNA and rDNA. Once again these deletions appear to be due to the presence of a duplicated region because all deletion products have lost one complete repeat. Deletions within both of these plasmids took place in both rec+ and recA- host cells, but occurred more frequently in rec+ cells. Oligomerization of the deletion products also occurred in both hosts and was more frequent in rec+ cells.

摘要

缺失发生在含有插入到大肠杆菌质粒pSC101和pMB9中的酵母核糖体DNA(rDNA)的重组DNA质粒中。在一个含有比一个串联rDNA重复序列更长插入片段的pMB9质粒内的缺失显然是由于同源重组,因为(1)该质粒所有独立衍生的缺失产物都丢失了一个完整的rDNA重复序列(8.6 kb),并且仅保留了在原始插入片段末端重复的片段的单拷贝,以及(2)仅当插入片段具有末端冗余时才检测到缺失。缺失也发生在一个含有包括pSC101 DNA和rDNA的片段(4.7 kb)串联重复的pSC101质粒内。这些缺失似乎再次是由于存在重复区域,因为所有缺失产物都丢失了一个完整的重复序列。这两种质粒内的缺失在rec⁺和recA⁻宿主细胞中均会发生,但在rec⁺细胞中发生得更频繁。缺失产物的寡聚化在两种宿主中也会发生,并且在rec⁺细胞中更频繁。

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