Cloning And Analysis Of Malespecific Expression Gene In Asparagus Officinalis

The animal is divided into male and female,Plants are also divided into female and male pla nts. The sex determination of most plants are XY. Dioecious plants is an excellent material to stu dy plant sex determination. Until now the key genes in sex determination and sex differentiation i n plants has not been foundAsparagus officinalis L. is aimportant perennial crops, typical of the dioecious plants. At present,it has been cloned somegenesrelated to flower development in asparagus,such as AODEF、AOGLOA、AOGLOB.In this paper,asparagus(UC309)maleandfemale plantsas materials, using semi-qu antitativeandreal-time quantitative technology analysis tissue-specific expressionin differencesgene. Cl oning of differentially expressed genes in male asparagus.The mainfindings are as follows:1. AO4-5 full-length genes equence was amplified by RACE technology, after the splices equ ences preliminary analysis showed that:AO4-5full length cDNA gene is 1348 bp, 5’UTR zone length of 77 bp, 3’UTR region length of 194 bp,which contains a full-length sequence of length 1077 bp O RF(open reading frame),the reading frame encoding 358 amino acid polypeptide chains.The amino acidsequence of AO4-5 alignment by NCBI Blastx online tools and in other plants found high a mino acid sequence homology, phylogenetictree was constructedthan the latter show that AO4-5 lik e protein similarity between the gene 43% to 66%; the highest similarity is Musa acuminate,the s imilarityof 66%.2. Use ExPasy, NetOglyx, NetNglyc, Coils Output, TMHMM,Signal P software AO4-5 gene for rbioinformatics analysis show that: The amino acid protein gene expression relative molecularw eightof39326.6, an isoelectric point(p I) 6.24; The polypeptide chain hydrophilic amino acids and hy drophobic amino acids uniform distribution in the peptide chain and hydrophobic index was-0.160,belonging to a hydrophilic protein;Thesecondary structure ofthe amino acidsequenceofthe genefor t heα-helix, β- irregularfolding andcurling, the proportion was33.80%, 13.97%and52.23%;The amino a cidsequence ofa total of13 phosphorylation sites, of whichfiveserine(Ser)site, athreonine(Thr)sites, seventyrosine(Tyr)sites; theamino acid sequence ofthe first seventoThe first29 amino acidslocated inthe t ransmembraneregion; SignalP analysis showedAO4-5gene hassignal peptideandsecretedproteins.3.ByRT-PCR, Q-PCRgeneexpression patternsofAO4-5analysis showed inmale and female aspara gus.AO4-5 gene only in the male expression. The expression level in male flower buds, male root,male stem, male leaf and female flower buds, female, female root, female stem, female leaf was basically identicalandfar less thaninthe expression ofthemale.AO4-5genethere is a cleardifferencein th emale and femaleorgans ofthe flowerpattern of expression, inother matureorganization does notexists ignificant differences.4. Speculate the AO4-5 gene protein expression of GDSL lipase,GDSLlipasebelong tohydrolas e familymembers. GDSL lipase is consist of serine,aspartateandhistidine and these triplets having ca talytic effect.Because of GDSLfamily ofserinelipase activityatthe N-terminus, soGDSLlipasehas beenc lassified asmembers of theSGNHhydrolase.In plants,a number of GDSL lipases have been cloned,Such as themodel plant Arabidopsis、canola、sorghum、corn and the others.Studies have shown tha t GDSL lipase is likely to play a key role in the growth and development of the regulatory mech anism of the plant.