1. 研究目的与意义
Content: Molecular cloning of the novel biosynthetic precursor-encoding cDNA from an Androctonus bicolor venom-derived cDNA library for the determination of DNA sequence and examination of gene expression.Meaning: Natural venom peptide templates are potential in the generation of analogues with enhanced biological activity and spectrum of actions that may be useful leads in the development of novel therapeutics.
2. 文献综述
Diverse methods in functional genomics studyYing Yuan( Faculty of pharmacy, Nanjing University of Chinese Medicine, Nanjing, China )Abstract:With the accomplishment of genome sequencing of rice and the launch of Human Genome Project (HGP), genomics research has entered the post genomics era, which means its focal point has been transferred from structural genomics area to functional genomics area. At the same time, the object of study changes from single genes to all genes. In other words, structural genomics represents the early stage of genomics analysis and is based on high-resolution genetic, physical and transcriptional mapping, providing the vast wealth of DNA and protein data, while functional genomics is a symbol of new era and characterized by high-throughput, large-scale experimental approaches (rather than a more traditional gene-by-gene approach), statistical analysis and computer analysis, using the database created by structural genomics to research into genes function. This article reviews the content and methods of functional genomics, including: microarrays, SAGE (serial analysis of gene expression), MicroRNA sequencing (miRNA-seq), functional protemics, bioinformatics. Together these measurement modalities endeavor to quantitate the various biological processes and improve our understanding of gene and protein functions and interactions.Key words:Functional genomics, content and methods, microarrays, SAGE, functional protemics.摘要：随着水稻基因组测序工作的完成以及人类基因组计划（HGP）的顺利进行，基因组学的研究已经由结构基因组研究为主转向进入了以功能基因组研究为标志的后基因组时代，其研究对象也由单一基因转变为大规模、批量分析。
换句话说，结构基因组学代表基因组分析的早期阶段，以建立生物体高分辩率的遗传、物理和转录图谱为主，提供了大量的DNA和蛋白质数据；功能基因组学代表基因分析的新阶段，以高通量、大规模实验方法及统计与计算机分析为特征，利用结构基因组学提供的信息系统地研究基因功能。
本文综述了功能基因组学的研究内容和方法，主要包括：微阵列；基因表达序列分析（SAGE）; miRNA测序；功能蛋白组学；生物信息学。
3. 设计方案和技术路线
Molecular Cloning of the Novel Biosynthetic Precursor-Encoding cDNA from an Androctonus Bicolor venom-Derived cDNA Library.A 5 mg sample of Androctonus bicolor venom was dissolved in1 mL of cell lysis/mRNA protection buffer supplied by Dynal Biotech, UK, and polyadenylated mRNA was isolated by magnetic oligo-dT beads as described by the manufacturer (Dynal Biotech, UK). The isolated mRNA was subjected to 5- and 3- rapid amplification of cDNA ends (RACE) procedures to obtain full-length AMP biosynthetic precursor nucleic acid sequence data using a SMART-RACE kit (Clontech, UK) essentially as described by the manufacturer. Briefly, the 3-RACE reactions employed a nested universal (NUP) primer (supplied with the kit) and a sense primer that was complementary to the N-terminal amino acid sequence. The 3-RACE reactions were purified and cloned using a pGEM-T vector system (Promega Corporation) and sequenced using an ABI 3100 automated sequencer. The sequence data obtained from these 3-RACE products were used to design a specific antisense primer to a conserved site within the 3-nontranslated region of the transcript. 5-RACE was carried out using this specific primer in conjunction with the NUP RACE primer and resultant products were purified, cloned, and sequenced.技术路线见附件
4. 工作计划
TIME CONTENT PROGRESS RATE2022.03.03~03.08 Read articles regarding to the experiment and do the presentation completed2022.03.09~03.23 Do the experiment completed2022.03.24~04.04 Read relative articles and write the initiating report and the review paper completed2022.04.05~04.15 Finish the experiment To be completed2022.04.16~05.15 Analyze the experimental data and write the research paper To be completed
5. 难点与创新点
We employed transcriptomic approaches combined with mass spectrometry to dissect the venom compositions of Androctonus bicolor.
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