SUPPRESSION OF DNAE NONSENSE MUTATIONS BY PCBA1
SUPPRESSION OF DNAE NONSENSE MUTATIONS BY PCBA1
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DOI：
10.1128/jb.171.6.3139-3143.1989
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发表时间：
1989-06-01
影响因子：
3.2
通讯作者：
MOSES, RE
中科院分区：
文献类型：
--
作者：
MAKI, H;BRYAN, SK;MOSES, RE
DNA聚合酶III被认为是大肠杆菌所必需的复制酶。DNA聚合酶III全酶的合成亚单位(α。亚单位)由dna E基因编码。我们已经报道，如果DNA聚合酶I和pcbA1突变存在，那么在没有功能dna E基因产物的情况下，大肠杆菌细胞可以存活和生长。现有的dna E基因突变是有条件缺陷的耐热突变。我们在这里报道了通过使用温度敏感的抑制突变来构建dna E基因的无义突变，以允许在允许的温度(32度)下存活。c)。PcbA1突变的引入消除了对温度敏感的表型。我们通过免疫印迹证实了缺乏可检测到的α。43度亚基。C。
DNA polymerase III has been recognized as the required replication enzyme in Escherichia coli. The synthesis subunit of DNA polymerase III holoenzyme (.alpha. subunit) is encoded by the dnaE gene. We have reported that E. coli cells can survive and grow in the absence of a functional dnaE gene product if DNA polymerase I and the pcbA1 mutation are present. Existing mutations in the dnaE gene have been conditionally defective thermolabile mutations. We report here construction of nonsense mutations in the dnaE gene by use of a temperature-sensitive suppressor mutation to permit survival at the permissive temperature (32.degree. C). Introduction of the pcbA1 mutation eliminated the temperature-sensitive phenotype. We confirmed by immunoblotting the lack of detectable .alpha. subunit at 43.degree. C.