Whole-sequencecdna Cloningand Exprission AnalysisofaccB AndaccCGenefromAleurites Moluccana(Linn.) Willd

Aleuritesmoluccana is a kind of woody oil plant which belongs to Euphorbiaceae. Its seeds contain high proportion of oil, which makes Aleurites moluccana to be an important biomass energy plant.In this experiment, whole-sequences cDNA of accB gene and accC gene of acetyl coenzyme A carboxylase BCCP subunit and BC subunit were extracted by cloning.And the sequences were analyzed in NCBI bybioinformatics method. Doing the RT-PCR semi-quantitative analysis of the expression of these two genes ofAleurites moluccana seeds in different growth stages is to provide a reference forscreeningmolecular marker or researching related enzymes about fat synthesis。(1) By homology cloning method combined with technology of RACE (cDNA endsRapid Amplification),obtained the full length sequences of accB and accC gene, and deduced their amino acid sequence by Bioinformatics.The BLAST program was used on NCBI website and related sofeware for analyzing and comparing. The result as follow:obtained an1170bp sequence of whole cDNA of accB gene, ORFwas798bp. The homology in nucleotide sequence of GeneaccB with Vernicia fordiiand Jatropha curcas which already registered in the GenBank were as follow:86%and81%. And the homology in amino acid sequence wite BCCP subunit of Vernicia fordii, Jatropha curcas, Medicago truncatula, Glycine max, Persea americana and Gossypium hirsutum andwere:91%,83%,68%,64%,61%, and52%.Obtaining an1607bp sequence of whole sequence of cDNA of accC gene, which contained1119bp ORF.The homology in nucleotide sequence of GeneaccC with Vernicia fordii and Jatropha curcas which already registered in the GenBank were as follow:86%and81%. Andthe homology in amino acid sequence with BC subunit of Jatropha curcas, Gossypium hirsutum, Nicotiana tabacum, Arachis hypogaea, Glycine max and Arabidopsis thalianaandwere:96%,91%,89%,89%,88%, and83%.(2) Taking the seed total RNA as a template, high quality cDNA was obtainedby using reverse transcription kit for seed total RNA transcription. Taking cDNA as a template and Actin about Aleurites moluccana Housekeeping gene as internal reference, expression of geneaccBand gene accC was analysised from Aleurites moluccana at different developmental stages of the seeds by semi-quantitative RT-PCR. In the process of seed development, expression quantity of accB gene rise quickly since may,and then decline slightly in July.But the quantity is higher in July than May. expression quantity of accC gene continue to rise since May to June,and then Rise gently in July.But there were not great difference between June and July. The conclusion:There are certain relationship between seed oil content of Aleurites moluccana and the expression of accB gene and accC gene.The expression quantity of accC gene are higher than accB gene.So result showed that the effect ofaccC gene on Aleurites moluccanafat synthesis may be greater than accB gene.