目的:构建小鼠Parkin co-regulated gene(Pacrg)/GFP-p Fast Bac1重组杆状病毒载体,并在Sf9昆虫细胞中表达。方法:PCR扩增小鼠全长Pacrg编码c DNA序列,通过TA克隆、连接等方法将该基因插入到携带e GFP的供体质粒p Fast Bac1中,获得重组...目的:构建小鼠Parkin co-regulated gene(Pacrg)/GFP-p Fast Bac1重组杆状病毒载体,并在Sf9昆虫细胞中表达。方法:PCR扩增小鼠全长Pacrg编码c DNA序列,通过TA克隆、连接等方法将该基因插入到携带e GFP的供体质粒p Fast Bac1中,获得重组载体rp FBac-Pacrg-GFP,然后转化到DH10Bac宿主菌中。筛选获得的重组杆状病毒载体r Bacmid-Pacrg-e GFP经脂质体转染至Sf9昆虫细胞中,荧光显微镜及Western印迹检测分析重组蛋白。结果:经测序及酶切鉴定显示构建的Pacrg/GFP-p Fast Bac1重组杆状病毒载体正确,Western印迹结果显示PACRG/e GFP融合蛋白在Sf9昆虫细胞中表达且携带绿色荧光。结论:成功构建了Pacrg/GFP-p Fast Bac1重组杆状病毒载体,且该重组蛋白在Sf9昆虫细胞中大量表达,为进一步研究PACRG蛋白的结构及其在精子发生中的调控作用奠定了基础。展开更多
This article explains the basis and ideas of modern social biologic theory. The author lays emphasis on the following opinions. Firstly, it is biology organism rather than gene that exist and multiply in the environme...This article explains the basis and ideas of modern social biologic theory. The author lays emphasis on the following opinions. Firstly, it is biology organism rather than gene that exist and multiply in the environment, so gene egoism is an inadequate determinism concept in biology. Secondly, it is actually difficult to tell which is the determinant factor: heredity or environment; besides, conscious ideas and human morality are also two types of heredity genes. So replacing human historical method with biology evolution method is improper in social biology. Thirdly, with the concept of gene culture common evolution, epigenesis regards choice, existence and adaptability as the main factors of cultural development, which actually has denied the freedom of human will, and changed common evolution into pure gene heredity. This results in the main contradiction in the whole inner contradictions of social biology.展开更多
文摘目的:构建小鼠Parkin co-regulated gene(Pacrg)/GFP-p Fast Bac1重组杆状病毒载体,并在Sf9昆虫细胞中表达。方法:PCR扩增小鼠全长Pacrg编码c DNA序列,通过TA克隆、连接等方法将该基因插入到携带e GFP的供体质粒p Fast Bac1中,获得重组载体rp FBac-Pacrg-GFP,然后转化到DH10Bac宿主菌中。筛选获得的重组杆状病毒载体r Bacmid-Pacrg-e GFP经脂质体转染至Sf9昆虫细胞中,荧光显微镜及Western印迹检测分析重组蛋白。结果:经测序及酶切鉴定显示构建的Pacrg/GFP-p Fast Bac1重组杆状病毒载体正确,Western印迹结果显示PACRG/e GFP融合蛋白在Sf9昆虫细胞中表达且携带绿色荧光。结论:成功构建了Pacrg/GFP-p Fast Bac1重组杆状病毒载体,且该重组蛋白在Sf9昆虫细胞中大量表达,为进一步研究PACRG蛋白的结构及其在精子发生中的调控作用奠定了基础。
文摘This article explains the basis and ideas of modern social biologic theory. The author lays emphasis on the following opinions. Firstly, it is biology organism rather than gene that exist and multiply in the environment, so gene egoism is an inadequate determinism concept in biology. Secondly, it is actually difficult to tell which is the determinant factor: heredity or environment; besides, conscious ideas and human morality are also two types of heredity genes. So replacing human historical method with biology evolution method is improper in social biology. Thirdly, with the concept of gene culture common evolution, epigenesis regards choice, existence and adaptability as the main factors of cultural development, which actually has denied the freedom of human will, and changed common evolution into pure gene heredity. This results in the main contradiction in the whole inner contradictions of social biology.
