Regulation of Ina in Escherichia Coli (Biology)

通过LacZ操纵子融合，在大肠杆菌中分离到一个新的pH依赖的基因座ina，定位时间为48.5min。该基因座受膜透性弱酸和原生质团诱导，但不受外部酸化的影响。这些数据可以用INA检测到的内部酸化来解释。将对INA融合进行克隆和测序，并对序列进行启动子和调控位点的分析。将通过克隆融合基因的体外转录来测试调控DNA的酸敏感性。H-回文酶在低pH值下产生H型DNA，其可能的调节作用将被研究。将通过为构成或不可诱导的分离株筛选Tn10插入和乙基甲烷磺酸盐突变体来分离INA的调节基因。互动活动包括教授一门分子生物学课程，举办一场关于妇女和科学主题的公开讲座，并为试图平衡竞争激烈的研究事业和家庭生活的年轻科学家组成一个支持小组。该项目进一步推动了VPW计划的目标，即：(1)为妇女提供机会，在NSF资助的工程和科学学科中发展她们的职业生涯；(2)通过提高受雇于工业、政府和学术机构的女科学家和工程师的知名度，鼓励妇女追求科学和工程事业。通过鼓励妇女参与科学，这是对国家未来科学活力的宝贵投资。

A novel pH-dependent locus ina, mapping at 48.5 min, has been isolated through lacZ operon fusions in Escherichia coli. This locus exhibits pH-dependent induction by membrane-permeable weak acids and by protonophores, but not by external acidification alone. These data could be explained by the detection of internal acidification by ina. The ina fusions will be cloned and sequenced, and the sequence analyzed for promoters and regulatory sites. Acid-sensitivity of the regulatory DNA will be tested by in vitro transcription of the cloned fusions. The possible regulatory role of H-palindromes, which produce H-form DNA at low pH, will be investigated. Regulator genes for ina will be isolated, by screening Tn10 insertions and ethylmethanesulfonate mutants for isolates which are either constitutive or uninducible. Interactive activities include teaching a course in molecular biology, presenting a public lecture on the topic of women and science, and forming a support group for young scientists attempting to balance competitive research careers with family life. This project furthers VPW program objectives which are (1) to provide opportunities for women to advance their careers in engineering and in the disciplines of science supported by NSF and (2) to encourage women to pursue careers in science and engineering by providing greater visibility for women scientists and engineers employed in industry, government, and academic institutions. By encouraging the participation of women in science, it is a valuable investment in the Nation's future scientific vitality.