Cloning And Expression Of KasⅢ Gene In Vernicia Fordii

Vernicia fordii is a great Chinese oil trees, is listed as one of China’s four major oil, Vemicia fordii seed oil that has been pressed with Tung oil, it is an important raw material。 KASⅢ is a kind of acyl carrier protein, It as in the synthesis of fatty acids fatty acyl carrier, It is started the key enzymes of fatty acid synthesis. It mainly work in a condensation reaction of fatty acid synthesis the first loop, cloning and researching expression of vernicia fordii KAS Ⅲ, it is important to reveal the molecular mechanism of Tung oil content. The paper primarily cloned KAS Ⅲ gene cDNA and genomic sequences of vernicia fordii, Studied the KAS ⅲ gene expression patterns, builded fordii KAS Ⅲ gene expression vector, Major findings are as follows:1.vernicia fordii KASⅢ gene cloning full length cDNA and genomic sequences (GenBank ID:1729461). Jane and degenerate fragment amplification526bp, Fragments of respectively870bp,630bp by5’RACE and3’RACE, Three fragments mosaics, get1complete cDNA sequence and length of1901bp. At the5’end UTR and3’end UTR of design primer amplification of cDNA full coding regions, respectively,sequencing results show that the fragment length of1651bp. The cDNA encoding box containing11209bp, coded403amino acids,3’5’ non-encoded non-coding region length is202bp and445bp, Found AATAA tail signals at1841bp,and signal19bp appeared with17poly-A tail, these suggest that amplification of aleurites fordii KASⅢ full length gene sequences.2.According to the cDNA sequence design primer, aleurites fordii genomic DNA as substrate amplification be of length5368bp genome sequences. Sequence in intron predicted the site, Analysis shows KASⅢ gene divided into8segments by7exons. Result of matching,each intron have the same length and location of the Castor and Jatropha.3.vernicia fordii KAS Ⅱ, KAS Ⅲ gene’s expression analysis:Analysis expression differences of different periods Vernicia fordii seed development and different organs and tissues. The expression of KASⅡgradually increased from June to September, at the end of September reached the maximum and then reduce; The expression of KAS Ⅲ gradually increased from June to August, late August reaches its maximum size, then gradually reduced in the different periods of Vernicia fordii seed development. In the different organs and tissues of vernicia fordii,the maximum expression of KAS Ⅱis in column headers,ovary is the second, the expression of KAS Ⅱ in mature tissue is more than it in immature tissue by analysis; The maximum expression of KAS Ⅲ is in delicate stems,the expression of KAS Ⅲ in immature tissue is more than it in mature tissue by analysis.4. vernicia fordii KAS Ⅲ gene expression vector construction:Build vernicia fordii KAS Ⅲ gene eukaryotic and prokaryotic expression vector by PCAMBIA1304and PET-30.