摘要
青霉素酰化酶基因(pac)的定位研究表明它的调节基因和结构基因位于3.5kb的Hind Ⅲ-EcoR Ⅰ片段。本文报道构建一系列质粒pPA 6的缺失衍生物,并测定这些缺失对pac表达的影响。结果表明调节基因位于pac结构基因内的Sph Ⅰ-Pvu Ⅱ片段。Sph Ⅰ-Pvu Ⅱ片段克隆到质粒pTZ18U,所得质粒pPA57转化到青霉素酰化酶产生菌E.coli D816,观察Sph Ⅰ-Pvu Ⅱ片段对染色体pac基因表达的影响。结果表明在Sph Ⅰ-Pvu Ⅱ片段内的调节基因反式调节pac基因表达。计算机分析表明在Sph Ⅰ-Pvu Ⅱ片段内有两个ORF是编码调节蛋白的可能候选者。pac调节基因的精确定位正在进行中。
The study on the localization of the pac gene indicated that the regulatory and structural genes were located in a 3.5 kb HindⅢ-EcoRI fragment. This paper reported that a series of deletions derivatives of plasmid pPA6 was constructed and the effect of deletions on the pac expression was determined. The results implied that the pac regulatory gene was located in the SphⅠ-PvuⅡ fragment within the pac structural gene. Then, the Sph Ⅰ-PvuⅡ fragment was cloned to the plasmid pTZ 18U at the Sph Ⅰ-Sma Ⅰ sites and the resulting plasmid pPA57 was transformed into the penicillin acylase producing strain E.coli D816 to observe the effect of Sph Ⅰ-PvuⅡ fragment on the chromosomal pac expression. The results demonstrated that the regulatory gene within the Sph Ⅰ-PvuⅡ fragment regulated the pac expression in trans. Computer analysis indicated that there were two ORFs in the Sph Ⅰ-PvuⅡ fragment as the possible candidates coding for the regulatory protein. The precise localization of the pac regulatory gene is underway.
出处
《生物工程学报》
CAS
CSCD
北大核心
1992年第2期105-115,共11页
Chinese Journal of Biotechnology
