軟件ssrmining1.0的編制以及大豆(glycinemax)estssr的開發(fā)研究

1、東北農(nóng)業(yè)大學(xué)碩士學(xué)位論文軟件SSRMINING1.0的編制以及大豆（Glycine max）EST-SSR的開發(fā)研究姓名：孫佳瑩申請學(xué)位級別：碩士專業(yè)：作物遺傳育種指導(dǎo)教師：陳慶山20090620ABSTRACT III Program of SSR MINING 1.0 and Identification and Characterization of EST-SSRs of Soybean (Glycine max) ABSTRA

2、CTEST-SSR is a type of molecular marker developed from the databank of sequence of expression sequence tags(EST)and/or cDNAs.As a new kind of molecular marker, EST-SSR markers have more advantages than the traditional ge

3、nomic-derived SSRs because they are part of expressed genes. Therefore, EST-SSRs might be involved in gene functioning directly or indirectly.The ability of SSR markers will be greatly enhanced. In this study, an efficie

4、nt arithmetic had been put forward by the analysis of the features of SSR marker itself and permutation and combination theory and corresponding software with Visual Basic 6.0 were developed. Then, primers based on the m

5、ining results from the nonredundant EST database in PlantGDB of soybean were designed. The distribution of SSR, design primers, filtered valid EST-SSR markers were analyzed with 8 soybean cultivars and part of these EST-

6、SSRs were mapped on the scaffold and linkage group of recent published soybean genome. Using BLAST, partial EST sequences were related to specific biological functions. 1. After summarizing and anglicizing the problems

7、of existed SSR markers developing software, a program SSR MINING 1.0 were accomplished to with Visual Basic 6.0. Then new SSR markers were searched from 22161 soybean EST sequences published between 2004. 08. 02 and 2005

8、. 06. 12 and 2070 SSR markers were detected, and with the help of DNAMAN, 30 pairs of primer had been designed for 30 SSR markers among them, then synthesized and verified by the experiments, 14 polymorphic SSR markers w

9、ere got. SSR MINING 1.0 is an efficient, flexible, interface friendly, and easy to use SSR markers developing software. 2. 200516 nonredundant soybean ESTs were found from the database PlantGDB, till April 5th, 2006. 12,

10、833 SSRs were discovered by the module TROLL that could insert in staden package. There was one SSR in every 7.25Kb EST in 93.1Mb soybean EST database. The frequencies of di-, tri-, tetra-, penta- SSR motifs and compoud

11、SSR were 25.2%, 46.5%, 6.9%, 16.1%, and 5.3% of total number of discovered SSRs respectly. 3. According to the selected EST-SSR sequences, primers were successfully designed from 9638 discovered EST-SSRs. 124 pairs prime

12、rs of these EST-SSRs were selected and synthesised. With genomic DNA of 8 soybean cultivars which represent a wide-range of diversity as template, PCR products of 81 pairs were successfully amplified while 44 of them sho

13、wed polymorphism, and could be used as new EST-SSR markers of soybean and related species. 4. The ESTs sequences corresponding to EST-SSR markers were analyzed by the programs BLAST of NCBI and Soybase to get the useful

14、information of possible function and expression pattern etc. After sequence similarity assignment, 29 ESTs have homologous sequences or proteins, and the locations were determined of 14 EST-SSR in genes. 5. 44 EST-SSRs w