ANALYSIS OF A BACTERIAL HYGROMYCIN-B RESISTANCE GENE BY TRANSCRIPTIONAL AND TRANSLATIONAL FUSIONS AND BY DNA SEQUENCING
ANALYSIS OF A BACTERIAL HYGROMYCIN-B RESISTANCE GENE BY TRANSCRIPTIONAL AND TRANSLATIONAL FUSIONS AND BY DNA SEQUENCING
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DOI：
10.1093/nar/11.19.6895
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发表时间：
1983-01-01
影响因子：
14.9
通讯作者：
INGOLIA, TD
中科院分区：
文献类型：
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作者：
KASTER, KR;BURGETT, SG;INGOLIA, TD
我们已经鉴定了来自 anE 的潮霉素 B 和安普霉素抗性基因。大肠杆菌质粒。我们通过从含有这些基因的质粒中删除DNA片段来定位这些基因的编码和控制区域。发现表观分子量为33,000和31,500道尔顿的多肽由安普霉素抗性基因编码，表观分子量为42,500和41,500道尔顿的多肽由潮霉素B抗性基因编码。 DNA 序列分析确定了基因上游的典型启动子序列。删除该启动子消除了两种抗性表型，并且可以通过替换外源基因的启动子来恢复潮霉素 B 抗性。已知潮霉素B抗性所必需的区域包含足够大的开放阅读框以编码潮霉素B抗性基因产物。该开放阅读框与 β-半乳糖苷酶的氨基末端融合。该杂交基因赋予大肠杆菌潮霉素抗性。大肠杆菌，耐药性的表达受到 IPTG 的控制。
We have characterized hygromycin B and apramycin resistance genes from anE. coliplasmid. 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 β-galactosidase. This hybrid gene conferred hygromycin resistance toE. coli, and expression of resistance was under IPTG control.