Construction Of Corchorus Capsularis Jute Blade Full-length CDNA Library And ESTs Analysis

Jute belonging to the Corchorus, Tiliaceae, is an annual herbaceous plant. So far, the function of jute gene research is still in the initial stage, it is difficult to draw the homologous sequences of other plant to study the functional genes. Therefore, the construction of jute full-length cDNA library can not only lay the foundation for juterelated genes and molecular biology studies,but also study the jute during the development of gene expression by the library screening,sequencing, and lay the foundation for a comprehensive understanding of the jute during the development of gene expression regulation.In this study, the leaves of the Capsularis jute 179 was materials, using SMART building a database technology to build jute leaf length cDNA library, and random parted on cloning sequencing,then, re-used bioinformatics processing and analysis. On this basis, the use of the EST sequence of development of jute-specific EST-SSR primers, it will also help the future of jute varieties clustering, the analysis of genetic diversity, genetic linkage map construction and other aspects of research. The specific research results were as follows:1. Compared the SDS method, CTAB method, hot boric acid-proteinase K method, Trizol method, the general plant kit and polysaccharide polyphenols kit six methods of extraction total RNA effect, then, by agarose gel electrophoresis and UV spectrophotometer detection of RNA integrity and purity, and the results showed that the effect of SDS method and general plant kit extraction of total RNA is ideal to meet the requirements of the follow-up experiments;2. The Corchorus capsularis Jute blade full-length cDNA library was constructed successfully, and the library qualification evaluation showed:the titer of primary cDNA library was 2.32×107 (pfu/mL), the titer of amplied library was 1.07×109 (pfu/mL). Randomly picked 48 monoclonal bacteria by PCR recombinants and found that the percentage of recombination was about 92%. In addition, the insert size concentrated between 500bp-1500 bp, with an average length of approximately 750bp; These results could be satisfied for further studying;3. Randomly picked 300 monoclonal bi-directional sequencing, success get 279 EST sequence, the success rate of 93%. Which low-quality ESTs were 63, high-quality ESTs were 216. The effective sequence length of these 216 ESTs span from 106bp-1386bp, all effective sequence the total length of 114820bp, the average length of 531.57bp, mean GC of 41.59%.61 unigenes of these 216 ESTs were spliced by using phrad Software: 24 contigs (including two or more than two ESTs) and 37 singlets (containing an EST sequence);4. With the software for the mutual repeat sequences comparison, statistics showed that jute leaves high abundance and mediem abundance of expressed genes in lower proportion (a total of 39.35%), but low abundance of expressed genes in the proportion of more than half (60.65%); Using BLASTX procedures, according to similar sequence of genetic structure and function description, compared with the non-redundant protein database (non-redundant protein database,nr library), after the functional annotation, found that there are 47 unigenes successfully compared with the previous and similar sequences total from 19 species; In addition, the GO classification also showed that there are 19 genes have been classified in the molecular function.