题目

Many young people want to become pop-stars. It is their great ambition（雄心）of life. They think: pop stars earn a lot of money; they lead interesting and exciting lives; thousands of fans love them; they become famous! You, but first they must become stars. Most performers start life with a group. This is the pattern nowadays. But there are many groups and the competition is hard. Groups appear and disappear almost overnight. Only a small number survive. Almost without exception（例外）they have to work very hard before they reach the top. And very few reach the top and stay there. I1994年，科研人员发现一株拟南芥突变株，由于NPR1基因突变，使其系统获得性抗性相关蛋白基因的应答性表达被彻底破坏，后来采用定位克隆技术从拟南芥中克隆到了NPR1基因，发现它的表达与植物抗细菌和真菌毒性提高有关。以NPR1基因作为目的基因可人工获得抗性植株。请回答：（1）抗性相关蛋白基因的应答性表达过程包括____________两个步骤。从野生拟南芥中分离出NPR1基因的mRNA后，以其为模板可_____________出目的片段，然后用PCR技术可实现目的片段的克隆。与生物体内的DNA复制相比，PCR技术的特殊之处有_______________(答出两项）等。（2）由于DNA复制时，子链只能由5＇向3＇方向延伸，故利用PCR扩增NPR1基因时，可以从图1的A、B、C、D四种单链DNA片段中选取___________作为引物。PCR扩增进行50个循环后，理论上可以产生约为___________个DNA分子，将这些DNA分子与载体连接后储存在一个受体菌群中，这个受体菌群就叫做拟南芥的_____________，扩增获得的目的基因与拟南芥细胞中该基因碱基序列_______(填“相同”或“不同”）。（3）将人工合成的NPR1基因导入普通植物之前，需要构建__________。对NPR1基因片段和质粒（图2所示）进行酶切时，可选用限制酶的组合为___________或_____________。