Kaster K R, Burgett S G, Rao R N, Ingolia T D
Nucleic Acids Res. 1983 Oct 11;11(19):6895-911. doi: 10.1093/nar/11.19.6895.
We have characterized hygromycin B and apramycin resistance genes from an E. coli plasmid. We have localized the coding and control regions of these genes by deletion of DNA fragments from plasmids containing the genes. It was found that polypeptides with apparent molecular weights of 33,000 and 31,500 daltons are encoded by the apramycin resistance gene and polypeptides with apparent molecular weights of 42,500 and 41,500 daltons are encoded by the hygromycin B resistance gene. DNA sequence analysis identified a typical promoter sequence upstream of the genes. Deletion of this promoter eliminated both resistance phenotypes, and hygromycin B resistance could be restored by substitution of a promoter from a foreign gene. The region known to be necessary for hygromycin B resistance contained an open reading frame large enough to encode the hygromycin B resistance gene product. This open reading frame was fused with the amino terminus of beta-galactosidase. This hybrid gene conferred hygromycin resistance to E. coli, and expression of resistance was under IPTG control.
我们已对来自大肠杆菌质粒的潮霉素B和阿泊拉霉素抗性基因进行了表征。我们通过从含有这些基因的质粒中缺失DNA片段，对这些基因的编码区和控制区进行了定位。结果发现，阿泊拉霉素抗性基因编码表观分子量为33,000和31,500道尔顿的多肽，潮霉素B抗性基因编码表观分子量为42,500和41,500道尔顿的多肽。DNA序列分析确定了这些基因上游的一个典型启动子序列。删除该启动子消除了两种抗性表型，通过替换来自外源基因的启动子可恢复潮霉素B抗性。已知对潮霉素B抗性必需的区域包含一个足够大的开放阅读框，可编码潮霉素B抗性基因产物。这个开放阅读框与β-半乳糖苷酶的氨基末端融合。这个杂种基因赋予大肠杆菌潮霉素抗性，且抗性表达受异丙基-β-D-硫代半乳糖苷（IPTG）控制。
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