In vitro mutagenesis of a Bacteriophge self-splicing Intron

根据分子生物学的中心教条，遗传物质DNA中包含的信息通过一种名为RAN的中间分子传递给蛋白质。蛋白质直接控制细胞的大部分功能。虽然基因表达的整个过程已经被理解，但涉及基因表达的机制的细节需要澄清。例如，插入序列(或内含子)是基因内部的额外信息片段，不用于产生最终的基因产品。基因内含子的重要性仍有待确定。该项目正在研究细菌病毒T4的剪接要求，这是一种在基因表达过程中从RNA中移除内含子的过程。RNA剪接机制多种多样，但在所有情况下，核苷酸(RNA和DNA的化学亚基)的序列对剪接过程都很重要。%在这个项目中，内含子的核苷酸序列将通过化学诱变产生变化，并将评估这些变化对剪接和基因表达的影响。这项研究的总体目标是利用化学诱导的内含子核苷酸序列的变化来识别内含子中对剪接重要的特定核苷酸。这些信息将用于增强拼接机构的模型。

According to the central dogma of molecular biology, the information contained in the hereditary material, DNA, is passed to proteins via an intermediate molecule called RAN. Proteins direct the control of most cell functions. While the overall process of gene expression is understood, the details of the mechanisms involved in gene expression need clarification. For example, intervening sequences (or introns) are extra pieces of information inside genes that are not used to produce the final gene product. The significance of introns inside genes remains to be determined. This project is looking at requirements for splicing , in the bacterial virus T4, a process that removes introns from RNA during gene expression. RNA splicing mechanisms are varied, but in all cases, the sequence of nucleotides (chemical subunits of RNA and DNA) is important to the splicing process. %%% In this project, changes in the nucleotide sequence of the introns will be generated by chemical mutagenesis and the effect of these changes on splicing and gene expression will be evaluated. The overall goal of this study is to use chemically induced changes in the nucleotide sequences of introns to identify particular nucleotides in the introns important to splicing. This information will be used to enhance models for splicing mechanisms.