Construction Of CDNA Library Of SuiNong 14 In The Filling-time And Analysis Of Expressed Sequence Tags

Soybean is the main edible oil and protein. The Soybean Genome research could conduce to know the genes in the Genome and help to drive the crop molecular development. Understanding the Soybean diseases and characters related genes will increase the crop yields and quality by Expressed Sequence Tag (EST) analysis. From the cDNA library, could find new genes and soybean genes, increase the density of the genetic maps, analyze the important functional genes and explain the gene expressed at the special tissue or development level.The soybean SuiNong14 is widely breed variety in our country. And it has much choiceness characters such as high content in oils and proteins. Constructing the cDNA library of SuiNong 14 in the filling-time and analyzing of Sequence are used to know the variety's characters. This research is get some important results as the following:1.Constructing the two cDNA library of seed under normal growth filling-time R5 and R6 respectively, were used by Creator SMART technique. In the primary library of R5 time, 1.2×106 clones were contained. PCR amplification demonstrated that the size of the inserted cDNA fragments ranged from 0.5 to 2kb, the average is 1.0kb and the recombination is nearly 99%.2.Analyze of the 1679 unigene by BLAST alignment, there are 1566 unigene are homolog to known genes. In these genes, the 74% have been gone beyond 80 score. And the most high expressed genes is energy metabolism, protein synthesis and storage class. The signal transduction and intracellular traffic class genes are expressed little in contract.3.Comparative of protease inhibitor in the Gm-R6 and some other seed-related libraries, the content changed with the seed development and expressed more in the mature time. Clone the 4 Bowman-Birk inhibitor use PCR method and analysis of the sequence discovered that there have new allele variance in the SuiNong14. Phylogenetic analysis of some crop BBI sequence demonstrated the evolutional relations.4.Combined the download seed-related cDNA library, analysis of the sequence function suggested that there are some genes highly expressed in the seed development as following: Auxin down regulated, Kunitz trypsin inhibitor, Chlorophyll a/b-binding protein, Lipoxygenase, Cationic peroxidase 2, Metallothionein type, Ubiquitin. In our constructed libraries, there are 65 genes take part in the lipid metabolism and more than 19 genes involved in the storage protein process. In the beginning of seed development the energy metabolism and synthesis class genes expressed more than others, however, the storage protein genes expressed most in the seed maturation time.