The RNA-seq Of Safflower And The Obtaining Of Its Oil Bodies Full Length Gene

With the rapid development of Molecular biology and genetic engineering, the vegetable oil bodies on bio-engineering were more and more widely. As the application of seed oil bodies in genetic engineering was used the oil body binding protein to achieve, so the study of the vegetable oil body binding protein has been paid attention to by people. Binding proteins of sesame oil, rapeseed, rice and Arabidopsis species have carried out some research on medicinal plants, while the study of safflower protein was little known, the current application has made some achievements, and the theory was just started. Safflower oil body protein gene sequences, protein sequences and regulatory factors are still unknown. Which are not good for the further in-depth study of Safflower oil body that can be as a bioreactor and the development of safflower oil body protein function that can be as food, medicine and surfactant.Solexa sequencing technology has been developed rapidly in recent years that is a high-throughput sequencing technology. Comparing to the traditional Sanger sequencing, high throughput sequencing has with a wide degree of information coverage and a deep sequencing feature. In recent years, Solexa sequencing and transcriptome analysis technology are established the macro-analysis for gene expression, which have high information covering, low redundancy and accurate analysis. It can detect low expressed genes exist and do not have the genomics background to analyze the transcriptional expression profiles of new species. and it also has many other advantages.cDNA library is one of the most effective ways to obtain full-length cDNA gene. Through various methods (such as differential display, suppression subtractive hybridization, etc.) to directly obtain EST fragments that can be as a screening probe to screen the cDNA library, using colony SOUTHERN hybridization to screen a complete cDNA clone from cDNA library. Using bioinformatics analysis method to obtain gene full-length cDNA and protein sequences.In this paper, using Solexa sequencing and bioinformatics analysis technology to study the transcriptome of seeds, leaves and flowers of safflower, then constructing cDNA library to obtain gene full-length cDNA the results as follow:1.The obtained safflower Unigenes were 153,769, including seeds of 69121; leaf of 51814; and flowers of 100739.2.The Unigenes functional annotation:including the Gene Ontology (GO) functional annotation; COG functional annotation, pathway annotation and protein functional annotation, the results:65,536 Unigenes were matched to the nr database; 56,843 Unigenes were matched to the Swiss-Prot database; 35,056 Unigenes were matched to the KEGG database; 11,010 Unigenes were matched to the COG database.3.Obtained 7.5×106 clones, and the successful application of bioinformatics methods selected from the cDNA library of the full length of safflower protein gene sequence by constructing cDNA library.