Molecular cloning of Cytosolic Ascorbate Peroxidase 1 gene from tomato plant and its bioinformatics analysis开题报告

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1. 研究目的与意义、国内外研究现状（文献综述）

课题的意义、国内外研究进展、应用前景等（列出主要参考文献）

significance

the tomato plant is considered to be a model plant of the solanaceae plant family. the significance of this research is that characterization of cytosolic ascorbate peroxidase 1 gene of tomato will provide an understanding of the structure, the evolutionary relationship of apx1 and its ancestral relation with other plants cytosolic apx gene and the related genes.

2. 研究的基本内容和问题

研究的目标、内容和拟解决的关键问题

experimental purpose

the purpose of this research is to clone the gene that encodes apx1 from a tomato plant. use the bioinformatics approach to analyze the structure of the target gene, and through a phylogenic tree, analyze the evolutionary relationship between tomato apx1 and other plants cytosolic apx gene.

3. 研究的方法与方案

研究方法、技术路线、实验方案及可行性分析

research methods

before the experimental research, literature reading is required. for the literature review, retrieving information about molecular cloning, cytosolic ascorbate peroxidase 1 gene, bioinformatics, and phylogenetic tree analysis the broad database google scholar will be used. from this literature, i believe i will be able to find experimental methods and protocols of molecular cloning. for bioinformatics purposes, i will download the software and find websites for bioinformatics analysis. mega software will be used for the multiple alignments of the sequence of tomato apx1 clone, cytosolic apx of other plant species, and the construction of phylogenetic for the analysis of the evolutionary relationship. tutorials and manuals for mega software are required to better understand how to operate the software. the website i will use for apx1 clone nucleotides blast, and download other plants cytosolic apx sequence, is the national center of biotechnology information (ncbi). to begin the experiment, i will use the methods and protocols found in the literature. experimental methods include rna extraction for the isolation of rna from tomato plant, the rna containing the whole genome of tomato plant will be transcribed into complementary dna through the method of reverse transcription, polymerase reaction chain (pcr) will be used to amplify the target gene (cytosolic ascorbate peroxidase 1) using specific forward and reverse primer, ligation, transformation and eventually sequencing.

4. 研究创新点

Molecular cloning of Cytosolic Ascorbate Peroxidase 1 (APX1) gene is significant for the investigation of plant development and plant stress tolerance against various environmental stimuli.

5. 研究计划与进展

研究计划及预期进展

25 february 2019 – 03 march 2019 retrieve literature related to the proposed topic from google scholar database.

04 march 2019 - 17 march 2019read all the information retrieved and make a compilation of notes, combine with the information i have from the literature review.