A Project-Oriented Molecular Biology/Genetics Laboratory Incorporating New Technologies

爱荷华州立大学（Iowa State University）的生物科学本科教育正在经历一场巨大的变革。从1993-94学年开始，正在实施一项新的生物学核心课程，包括对爱荷华州立大学所有生命科学专业的两年综合教学。生物学核心课程第二年的一个关键组成部分将是遗传学和分子生物学实验课程，这将为本科生提供一个重要的机会，让他们在本科和以后的职业生涯中获得研究机会。我们为这个实验室设计了一系列具有挑战性的，以项目为导向的练习，包括突变特征的遗传方法和分子生物学中最先进的方法。该项目从分离受嘧啶代谢干扰的酵母菌突变体开始，重点鉴定尿嘧啶生物合成基因URA3的突变体。学生将通过互补分析表征URA3突变体，并使用分子方法克隆野生型URA3基因。首先将进行计算机练习，使学生熟悉在实验室中进行的分子操作。然后，学生们将通过聚合酶链反应从基因组DNA中扩增出URA3基因的野生型副本，将该基因克隆到可以在大肠杆菌和酵母中发挥作用的质粒载体中，并通过琼脂糖凝胶电泳和Southern杂交分析来表征推定的重组基因。在最后的练习中，学生们将通过将他们的URA3突变体转化为克隆的野生型URA3基因来补充他们。所要求的设备用于酵母的生长和转化，以及DNA扩增、纯化和分析。爱荷华州立大学将为该基金会提供同等数额的资金。

Undergraduate education in the biological sciences at Iowa State University is undergoing a dramatic change. Beginning in the 1993-94 academic year, a new core curriculum in biology is being implemented, consisting of two years of integrated instruction for all life sciences majors at Iowa State. A key component of the second year of the biology core will be a genetics and molecular biology laboratory course, which will provide an important opportunity to capture the interest of undergraduates for research opportunities both during their undergraduate careers and beyond. We have designed for this laboratory, a series of challenging, project-oriented exercises that encompass both genetic approaches to mutant characterization and the most advanced methods in molecular biology. The project begins with the isolation of yeast mutants perturbed in pyrimidine metabolism, focusing on the identification of mutants in the uracil biosynthetic gene, URA3. Students will characterize URA3 mutants by complementation analyses, and use molecular methods to clone a wild-type URA3 gene. A computer exercise will initially be conducted to familiarize students with the molecular manipulations carried out in the laboratory. Students will then proceed to amplify a wild-type copy of the URA3 gene from genomic DNA by the polymerase chain reaction, to clone this gene in a plasmid vector that can function both in E. coli and yeast, and to characterize putative recombinants by agarose gel electrophoresis and Southern hybridization analysis. In the final exercise, students will complement their URA3 mutants by transforming them with the cloned, wild-type URA3 gene. The equipment requested provides for growing and transforming yeast, as well as DNA amplification, purification and analysis. Iowa State University will provide an equal match for the funding from the foundation.