High-throughput Analysis of Brugia malayi mRNA 5' ends

马来丝虫 mRNA 5 末端的高通量分析

• 批准号: 6963272
• 负责人: Elodie Ghedin
• 金额: $ 0.77万
• 依托单位: INSTITUTE FOR GENOMIC RESEARCH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6963272
• 关键词: Wuchereria; genetic library; genetic transcription; high throughput technology; messenger RNA; method development; nucleic acid sequence; nucleic acid structure

Brugia malayi is a causative agent of lymphatic filariasis, a disease which affects 120 million individuals in tropical and subtropical regions of the globe. As the assembly of the 6. malayi genome nears completion, an outstanding task is the full prediction and structural annotation of its genes. A lack of experimental evidence and of full length cDNAs has limited the efficient training of gene finder algorithms. A critical marker in the assessment of gene structure is the determination of the 5' transcriptional start site of each gene. We propose to generate a comprehensive catalog of B. malayi 5' ends by using two novel methods. First, we propose to use a recently developed technique, Trans-spliced Exon Coupled RNA End Determination (TEC-RED), to make a library of concatenated 5' sequence tags representing all SL1 trans-spliced genes in B. malayi. Second, we propose a novel variation of the TEC-RED technique to characterize 5' transcriptional start sites using a modified oligo-capping methodology. In the course of developing this novel protocol we propose to construct and sequence an oligo capped library which will be enriched for complete capped mRNA sequences. The analysis of the 5' sequence of a significant portion of B. malayi mRNAs will provide a number of benefits for the annotation of the B. malayi genome, including determination of transcriptional initiation sites, alternative splice sites and identification of transcriptional start sites of previously unidentified genes. The compilation of B. malayi-full-length mRNAs will also aid in the discovery of novel splice leader sequences. This could potentially lead to the discovery of previously undiscovered operons in 8. malayi. Moreover, the collection of trans-spliced genes will serve as a comparative marker for evolutionary studies in nematodes.

马来丝虫是淋巴丝虫病的病原体，淋巴丝虫病是一种影响地球仪热带和亚热带地区1.2亿人的疾病。作为第六届大会。马来人基因组接近完成，一个突出的任务是对其基因进行全面预测和结构注释。缺乏实验证据和全长cDNA限制了基因查找算法的有效训练。基因结构评估中的关键标记是确定每个基因的5'转录起始位点。我们建议生成B的综合目录。Malayi 5'通过使用两种新颖的方法结束。首先，我们建议使用最近开发的技术，反式剪接外显子偶联的RNA末端测定（TEC-RED），使连接的5'序列标签库代表所有的SL 1反式剪接基因在B。马来语。其次，我们提出了一种新的变化的TEC-RED技术，以表征5'转录起始位点使用修改的寡加帽方法。在开发这种新方案的过程中，我们提出构建和测序寡核苷酸加帽文库，其将富集完整的加帽mRNA序列。B的重要部分的5'序列的分析。马来人的mRNA将为B的注释提供许多益处。马来人基因组，包括确定转录起始位点，选择性剪接位点和以前未鉴定的基因的转录起始位点的鉴定。B的编制。马来全长mRNA也将有助于发现新的剪接前导序列。这可能会导致在8中发现以前未发现的操纵子。马来语。此外，反式剪接基因的收集将作为线虫进化研究的比较标记。