摘要： G蛋白偶联受体普遍存在于生物体内，它是人类体内数量最大的细胞表面受体一族。由于细胞膜上这些聪明的受体，G蛋白偶联受体的存在，让我们发现了人体内的五彩缤纷。至今为止，人们已经发展研究出多种原核和真核的表达系统，这些系统被用来生产重组蛋白质。通过研究发现，经杆状病毒侵染的昆虫细胞是GPCRs最普遍的宿主，也一种比较通用的模型被用来研究GPCRs生理特性。而且昆虫细胞更利于培养，它的蛋白活性可比似哺乳细胞。49677

聚合酶链式反应（PCR）技术用于分子生物学中，可以在几个数量级范围内扩增一个单拷贝或者多拷贝的DNA片段，同时生成数千到数百万特定的DNA序列。这项技术对于扩增一个较为集中的DNA片段是既方便又廉价的，它在遗传疾病的诊断和监测，识别犯罪（在法医学领域），以及研究目标函数段等方面有着重要作用。

毕业论文关键词：GPCR受体，PCR实验，质粒提取，双切酶，SF9昆虫细胞，大肠杆菌。

The expression of Heterologous genes in insect cells

Abstract： G protein coupled receptor (GPCR)is ubiquitous  in the organisms, which is the largest family of cell surface receptor in the human body.Due to these clever receptor on the cell membrane――the Existence of GPCR,our body can feel this colorful world.So far, people have developed a variety of prokaryotic and eukaryotic expression systems to produce recombinant protein.Insect baculovirus infection is a commonly-used host cells,to study GPCR , and is also the general model to study the physiological characteristics of GPCR .insect cells are beneficial to mass culture and they have the protein activity which is simliar to mammalian cells.

The polymerase chain reaction (PCR) is a technique used in molecular biology to amplify a single copy or a few copies of a piece of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It is an easy and cheap tool to amplify a focused segment of DNA, useful in the diagnosis and monitoring of genetic diseases, identification of criminals (under the field of forensics), studying the function of targeted segment, etc.

Keywords：G protein coupled receptor (GPCR)，PCR，plasmid，Enzyme, insect cells, e. Coli.

目录

1  绪论 1

1.1  研究意义和目的 3

1.1.1 意义 3

1.1.2 目的 3

1.2  真核膜蛋白的重组表达系统 4

1.2.1大肠杆菌 4

1.2.2 酵母细胞 5

1.2.3哺乳动物细胞 5

1.2.4 昆虫细胞 6

1.3 GPCR在昆虫细胞中的表达 7

2  实验部分 8

2.1实验基本过程 8

2.2实验仪器及试剂 8

2.3实验操作步骤 9

2.3.1 PCR实验 9

2.3.2琼脂糖凝胶电泳 10

2.3.3 AxyPrep DNA凝胶回收试剂盒 10

2.3.4 限制性核酸内切酶 11

2.3.5 大肠杆菌DH5α感受态的制备和质粒的转化 11

2.3.6质粒摇菌培养 12

2.3.7 AxyPrep质粒DNA小量试剂盒