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一个用于强化基因结构与表达并分析基因与蛋白质序列之间关系的生物信息学入门练习。

An introductory bioinformatics exercise to reinforce gene structure and expression and analyze the relationship between gene and protein sequences.

作者信息

Almeida Craig A, Tardiff Daniel F, De Luca Jane P

机构信息

Biochemistry Program, Stonehill College, Easton, Massachusetts 02357; Biology Department, Stonehill College, Easton, Massachusetts 02357.

出版信息

Biochem Mol Biol Educ. 2004 Jul;32(4):239-45. doi: 10.1002/bmb.2004.494032040373.

Abstract

We have developed an introductory bioinformatics exercise for sophomore biology and biochemistry students that reinforces the understanding of the structure of a gene and the principles and events involved in its expression. In addition, the activity illustrates the severe effect mutations in a gene sequence can have on the protein product. Students search GenBank for the wild-type nucleotide sequence of the Caenorhabditis elegans unc-22 gene, the amino acid sequence of its gene product, and the nucleotide sequence of the transposon Tc5. The nucleotide sequences are manipulated using two programs in the Lasergene® software package from DNASTAR®. The first program, EditSeq®, enables students to experience the meticulous process required to precisely locate and remove intron sequences from the wild-type unc-22 allele to generate a cDNA sequence. The unc-22(r466) allele is generated by inserting the sequence of the transposon Tc5 into the appropriate location of the third exon in unc-22. The open reading frames of both cDNAs are located and then translated. MegAlign®, the second program, aligns the wild-type sequence of the UNC-22 protein and the wild-type and mutant protein sequences that were constructed. The degree of sequence similarity between the aligned proteins allows students to verify their success in processing the gene, as well as to visualize the truncated protein product from the Tc5 mutant allele. Student feedback and possible modifications to the exercise as well as supplemental exercises are also discussed.

摘要

我们为大二的生物学和生物化学专业学生开发了一项生物信息学入门练习,以强化他们对基因结构以及基因表达所涉及的原理和过程的理解。此外,该活动还展示了基因序列中的突变对蛋白质产物可能产生的严重影响。学生们在GenBank中搜索秀丽隐杆线虫unc-22基因的野生型核苷酸序列、其基因产物的氨基酸序列以及转座子Tc5的核苷酸序列。使用DNASTAR®公司的Lasergene®软件包中的两个程序对核苷酸序列进行操作。第一个程序EditSeq®,让学生体验从野生型unc-22等位基因中精确定位和去除内含子序列以生成cDNA序列所需的细致过程。unc-22(r466)等位基因是通过将转座子Tc5的序列插入unc-22第三个外显子的适当位置而产生的。确定两个cDNA的开放阅读框,然后进行翻译。第二个程序MegAlign®,将UNC-22蛋白的野生型序列与构建的野生型和突变型蛋白序列进行比对。比对后的蛋白质之间的序列相似程度使学生能够验证他们在处理基因方面的成功,同时也能直观看到来自Tc5突变等位基因的截短蛋白产物。还讨论了学生的反馈、对该练习可能的修改以及补充练习。

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