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酿酒酵母中细菌β-内酰胺酶基因的转录

Transcription of the bacterial beta-lactamase gene in Saccharomyces cerevisiae.

作者信息

Breunig K D, Mackedonski V, Hollenberg C P

出版信息

Gene. 1982 Nov;20(1):1-10. doi: 10.1016/0378-1119(82)90082-8.

Abstract

We have examined the transcription in yeast of Escherichia coli-yeast 2-micrometers DNA recombinant plasmids carrying the bacterial beta-lactamase (bla) gene. In Saccharomyces cerevisiae both strands of the gene are transcribed giving multiple RNA species of distinct lengths. At least one RNA transcript derived from the coding strand initiates at a yeast promoter on the 2-micrometers DNA segment. Another mRNA of 1.1 kb starts right in front of the gene on the bacterial DNA sequence. Deletion experiments have shown that expression of the bacterial bla gene is dependent on the presence of bacterial sequences right in front of the gene. Mutants lacking the bacterial promoter region do not give detectable gene products in yeast. The expression can be restored by substituting for the deleted sequence a DNA fragment which carries the E. coli lac promoter-operator region. We conclude that the bacterial promoter region of the bla gene as well as the lac promoter-operator fragment have promoter activity in yeast and that yeast-bla fusion transcripts cannot be used as a functional messenger for beta-lactamase in yeast.

摘要

我们研究了携带细菌β-内酰胺酶(bla)基因的大肠杆菌-酵母2微米DNA重组质粒在酵母中的转录情况。在酿酒酵母中,该基因的两条链都被转录,产生多种长度不同的RNA种类。至少有一种源自编码链的RNA转录本起始于2微米DNA片段上的酵母启动子。另一种1.1 kb的mRNA正好起始于细菌DNA序列上该基因的前方。缺失实验表明,细菌bla基因的表达依赖于该基因前方细菌序列的存在。缺乏细菌启动子区域的突变体在酵母中无法产生可检测到的基因产物。通过用携带大肠杆菌乳糖启动子-操纵子区域的DNA片段替代缺失序列,可以恢复表达。我们得出结论,bla基因的细菌启动子区域以及乳糖启动子-操纵子片段在酵母中具有启动子活性,并且酵母-bla融合转录本不能用作酵母中β-内酰胺酶的功能性信使。

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