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中国仓鼠卵巢细胞中二氢叶酸还原酶基因的结构

Structure of the dihydrofolate reductase gene in Chinese hamster ovary cells.

作者信息

Carothers A M, Urlaub G, Ellis N, Chasin L A

出版信息

Nucleic Acids Res. 1983 Apr 11;11(7):1997-2012. doi: 10.1093/nar/11.7.1997.

DOI:10.1093/nar/11.7.1997
PMID:6300788
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC325857/
Abstract

Overlapping recombinant lambda 1059 phages carrying regions of the dhfr locus from the amplified Chinese hamster ovary (CHO) cell clone MK42 have been isolated. In addition, dhfr cDNAs from this cell line have been cloned into plasmid pBR322. Restriction analysis of these recombinant molecules has led to a map of the Chinese hamster dhfr gene. This gene has a minimum size of 26 kb and contains six exons as defined by hybridization to a combination of mouse and CHO cDNA probes. The latter probes reveal 3' exonic sequences that are not present in mouse cDNA. The CHO dhfr gene thus extends about 700 bp further 3' than in the mouse, consistent with the larger size of the hamster mRNA. At least five intervening sequences are present, of approximate sizes: 0.3, 2.5, 8.6, 2.6 and 9.4 kb. Four sequences from highly repeated families are situated in introns within the dhfr gene. The overall structure of this gene is strikingly similar to that of the mouse. Evolutionary conservation of interrupted gene structure among mammals thus extends to genes that code for household enzymes as well as specialized or structural proteins.

摘要

已分离出携带来自扩增的中国仓鼠卵巢（CHO）细胞克隆MK42的二氢叶酸还原酶（dhfr）基因座区域的重叠重组λ1059噬菌体。此外，已将来自该细胞系的dhfr cDNA克隆到质粒pBR322中。对这些重组分子的限制性分析已得出中国仓鼠dhfr基因的图谱。该基因的最小大小为26 kb，包含六个外显子，这是通过与小鼠和CHO cDNA探针的组合杂交确定的。后一种探针揭示了小鼠cDNA中不存在的3'外显子序列。因此，CHO dhfr基因在3'端比小鼠的延伸约700 bp，这与仓鼠mRNA的较大尺寸一致。至少存在五个间隔序列，其大致大小分别为：0.3、2.5、8.6、2.6和9.4 kb。来自高度重复家族的四个序列位于dhfr基因内的内含子中。该基因的总体结构与小鼠的极为相似。因此，哺乳动物中中断基因结构的进化保守性延伸到编码常用酶以及特殊或结构蛋白的基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/f730e21c47c0/nar00352-0061-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/15db4aba0873/nar00352-0056-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/63f7dc616a84/nar00352-0056-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/5e7cada43ee3/nar00352-0057-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/310fb900ca9a/nar00352-0059-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/c56fad98917e/nar00352-0060-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/f730e21c47c0/nar00352-0061-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/15db4aba0873/nar00352-0056-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/63f7dc616a84/nar00352-0056-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/5e7cada43ee3/nar00352-0057-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/310fb900ca9a/nar00352-0059-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/c56fad98917e/nar00352-0060-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3da6/325857/f730e21c47c0/nar00352-0061-a.jpg

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