Transgenic safety issues cause more and more controversies with the planting area of transgenie crops increased year by year. Gene flow from transgenie crops to wild relatives through pollen dispersal is one of the fo...Transgenic safety issues cause more and more controversies with the planting area of transgenie crops increased year by year. Gene flow from transgenie crops to wild relatives through pollen dispersal is one of the focus problems. Gene splitting technique provides a new strategy for the control of transgene flow by bio-logical containment. The construction of gene splitting technique is based on protein trans-splicing mediated by intein. Currently, it has been proved in Arabidopsis, tabaoco, wheat, etc. that active and functional proteins can be reassembled by intein mediated protein trans-splicing after gene splitting, which provides theoretical basis and experimental supporting for the limit of transgene flow by gene splitting technique. The theoretical basis of gene splitting technique and research progresses of its application on the control of transgene flow were reviewed in this paper.展开更多
为建立高效快捷的蛛丝功能化修饰平台,蛋白质内含子的反式剪接技术被首次应用于重组蛛丝的功能化修饰。在体外通过Ssp Dna B的反式剪接作用,在蛋白质水平上将12 k Da泛素相关修饰蛋白(SUMO)与蛛丝蛋白(W2CT)连接形成功能化蛛丝蛋白SUMOW...为建立高效快捷的蛛丝功能化修饰平台,蛋白质内含子的反式剪接技术被首次应用于重组蛛丝的功能化修饰。在体外通过Ssp Dna B的反式剪接作用,在蛋白质水平上将12 k Da泛素相关修饰蛋白(SUMO)与蛛丝蛋白(W2CT)连接形成功能化蛛丝蛋白SUMOW2CT。修饰后SUMOW2CT与W2CT均能形成纳米至微米级的丝纤维,但SUMOW2CT自动成丝速度明显下降且产量约为W2CT的一半。与W2CT丝纤维(W)相似,SUMOW2CT丝纤维(UW)不具有超收缩能力和对2%SDS不耐受,但机械性能低于W2CT丝纤维。功能化蛋白SUMOW2CT形成的丝纤维中SUMO蛋白仍保持着正确三维结构,可被SUMO蛋白酶酶切。外源功能化蛋白质虽在一定程度上降低了丝的形成速度和机械性能,但修饰上的功能化蛋白仍保持着生物活性,表明断裂蛋白质内含子介导的蛛丝修饰平台成功建立,也为蛛丝的功能化修饰和应用奠定了坚实的技术基础。展开更多
基金Supported by Major Project of China on New Varieties of GMO Cultivation(2013zx08010-003)National Natural Science Foundation of China(31100408)
文摘Transgenic safety issues cause more and more controversies with the planting area of transgenie crops increased year by year. Gene flow from transgenie crops to wild relatives through pollen dispersal is one of the focus problems. Gene splitting technique provides a new strategy for the control of transgene flow by bio-logical containment. The construction of gene splitting technique is based on protein trans-splicing mediated by intein. Currently, it has been proved in Arabidopsis, tabaoco, wheat, etc. that active and functional proteins can be reassembled by intein mediated protein trans-splicing after gene splitting, which provides theoretical basis and experimental supporting for the limit of transgene flow by gene splitting technique. The theoretical basis of gene splitting technique and research progresses of its application on the control of transgene flow were reviewed in this paper.
基金Supported by National Natural Science Foundation of China(No.30800186 and 31070698)National High Technology Research and Development Program of China(863 Program,No.2006AA03Z451)+2 种基金Shanghai Key Project of Basic Program(No.10JC1400300)Ph.D. Programs Foundation of Ministry of Education of China(No.200802551026)Special Funds of Basic Scientific Research Operating Expenses from Chinese Universities(No.11D10530)~~
文摘为建立高效快捷的蛛丝功能化修饰平台,蛋白质内含子的反式剪接技术被首次应用于重组蛛丝的功能化修饰。在体外通过Ssp Dna B的反式剪接作用,在蛋白质水平上将12 k Da泛素相关修饰蛋白(SUMO)与蛛丝蛋白(W2CT)连接形成功能化蛛丝蛋白SUMOW2CT。修饰后SUMOW2CT与W2CT均能形成纳米至微米级的丝纤维,但SUMOW2CT自动成丝速度明显下降且产量约为W2CT的一半。与W2CT丝纤维(W)相似,SUMOW2CT丝纤维(UW)不具有超收缩能力和对2%SDS不耐受,但机械性能低于W2CT丝纤维。功能化蛋白SUMOW2CT形成的丝纤维中SUMO蛋白仍保持着正确三维结构,可被SUMO蛋白酶酶切。外源功能化蛋白质虽在一定程度上降低了丝的形成速度和机械性能,但修饰上的功能化蛋白仍保持着生物活性,表明断裂蛋白质内含子介导的蛛丝修饰平台成功建立,也为蛛丝的功能化修饰和应用奠定了坚实的技术基础。
