变形链球菌F-ATP酶基因重组质粒的构建和初步分析

目的:构建用于转化变形链球菌,含有F-ATP酶基因的重组质粒。方法:采用PCR方法,以变形链球菌基因组DNA为模板,扩增F-ATP酶β亚基5′末端序列,将克隆片段与载体pVA891酶切后连接,形成重组质粒,并对变形链球菌的转化作了初步分析。结果:构建的重组质粒经PCR鉴定、酶切鉴定和DNA序列测定,显示插入的目的片段序列正确。结论:变形链球菌目的片段与穿梭载体重组后能有效克隆,为通过同源重组特异突变变形链球菌染色体基因奠定基础。

Identification of Myosinon the Surface of Wheat Mitochondria

Itwasdemonstratedthatmyosinwasassociatedwiththesurfaceofmitochondriainwheat (Triticum aestivumL .) .AssaysofpolyacrylamidegelelectrophoresisandWesternblottinghaveshownthatapolypeptide withmolecularweightof2 10kDcouldberecognizedbyapolyclonalantibodyagainsthumanmusclemyosin .It wasfoundthattheATPaseactivityofmitochondrialsuspensioncouldbestimulatedbyF_actinisolatedfrom chickenmuscle ,whichindicatedthattherewasmyosinonthesurfaceofwheatmitochondria .Thisresultwas confirmedbyelectronmicroscopicobservation :mitochondriatreatedwithN_ethylmaleimide (NEM)couldbe wrappedbytheF_actin .