The Hetheroplasmy Of Chicken Mitochondrial ND2Gene And The Effect Of Calorie Restriction And Oil Source On The Hetheroplasmy

ND2gene is a protein-coding genes of mitochondrial DNA, constitutes one ofthe important subunits of mitochondrial respiratory chain complexes，and participatein mitochondrial respiratory chain and oxidative phosphorylation function, expoundsthe energy plays a very important role. In preliminary tests, we found that the ND2gene contains5703A> T,5727T> G, a lot of potential sites of heteroplasmy, So wespeculated that the chicken mtDNA genome heteroplasmy is likely to be a widespreadphenomenon.To explore what factors influence the chicken mitochondria ND2geneheteroplasmy, this study adopted the pyrophosphate sequencing technology, examinedthe heteroplasmy of the different organizations,30%energy limit the organization offeeding group and different oil added group organization. The results show that thereare considerable differences in their heteroplasmy of mtND25703A> T and5727T> G between twelve groups in the same individual. MtND2locus on5727T> G, thechest (the percentage bases G is96.9%), leg muscle ((the percentage bases G is96.4%), bursa of fabricius ((the percentage bases G is54.2%);And, mtND25703A>T, chest muscle tissue (the percentage bases T is93%), leg muscle (the percentagebases T is94.8%), bursa of fabricius (the percentage bases T is47.8%). However,including the brain, the cerebellum, small intestine, kidney, lung, liver, glandularstomach, muscular stomach and heart, these organizations in the two sites have noheteroplasmy. This means that the chicken mitochondria heteroplasmy and its degreehas obvious difference in the organizations, and in the chest, leg muscle, bursa offabricius organization, has higher heteroplasmy.We studied the effect of30%energy limit on chicken heterogeneity,and thepyrophophate sequencing results showed that mtND25727T> G site, The30%energy restricted and freed feeding groups are TT homozygous type (base Tpercentage is100%), Indicates that mtND25727T> G sites, the chicken breastmuscle of two groups have no heteroplasmy; MtND25703A> T sites, The30%energy restricted and freed feeding groups are AA homozygous type (base Apercentage is100%), Show that experimental chicken chest mtND25703A> T locihave no heterogeneity. This also means that the diet30%of the energy limitation,does not affect the chicken ND2gene mtND2mtND25703A> T and5727T> Glocus heteroplasmy in the chest.Pyrophosphate sequencing method is used to study the heteroplasmy of legmuscle groups which foods were adding5%linseed oil,5%lard,5%sesame oil,5%corn oil.Results:5727T> G mtND2loci, four oil group is TT homozygous type(base T percentage is100%),Show that experimental chicken chest mtND25727T> G sites have no heteroplasmy; MtND25703A> T loci, four oil group are AAhomozygous type (allele A percentage of100%) showed that experimental chickenchest mtND25703A> T loci were not heterogeneity. This also means that differentoil levels did not cause ND2gene mtND2mtND25703A> T and5727T> G loci inthe heteroplasmy of chest muscle and its degree of heteroplasmy.Using high fidelity Taq polymerase to cloning sequencing of six individuals, wedetected29mutation loci on642bp ND2gene sequences,Including mtND25727T>G,5703A> T two loci,Altogether constitute the21kinds of haploid type,Shows themitochondrial gene mutation of generalized potential richness and heterogeneous.This study explores the, influence of energy restriction and different oils on themitochondrial heteroplasmy, And find that the chicken mitochondria heteroplasmyand its degree has obvious difference in different organizations. Cloning sequencingfound multiple mutation loci, shows the mitochondrial gene mutation of generalizedpotential richness and heterogeneous.It provides an important reference onheteroplasmy of mitochondria for further research.