Title Phosphoprotein prokaryotic and mass spectrometric

analysis of muscle sarcoplasmic reticulum by people。

Abstract Human muscle sarcoplasmic reticulum by phosphoprotein, cardiomyocytes (PLN) is a membrane protein, composed of 52 amino acid, can in the myocardium and skeletal muscle directly inhibits muscle sarcoplasmic reticulum calcium pump -Ca2+-ATPase (SERCA)。 Phospholamban (PLN) through reduction in SERCA and Ca2+-binding, thereby weakening of SERCA function。 However, phosphorylation by phosphoprotein can reduce the inhibition of SERCA。 But interacting proteins in the molecular mechanism of protein so far is not clear, therefore phospholamban polymerization state, phosphorylation and binding to the membrane lipid molecules, has been the focus of people's attention。 Constructs containing the HB tag by phospholamban (PLN) expression vector, this experiment using molecular cloning method and transformed into Escherichia coli expression strain BL21 (DE3), try for the expression and purification of phosphoprotein PLN, in order to obtain high purity mg of PLN, PLN to use the mechanism of mass spectrometry analysis lay the foundation。

Keywords  Molecular cloning  molecular mechanism  phospholamban protein-expression

目 录

1 绪论 1

1。1 受磷蛋白概述 1

1。2 受磷蛋白的结构特点 1

1。3 受磷蛋白的功能特性 2

1。4 受磷蛋白磷酸化作用 3

1。5 受磷蛋白的研究及其意义 3

1。6 受磷蛋白 4

1。7 基因克隆 5

1。7。1 基因克隆技术 5

1。7。2 基因克隆技术的发展历程 6

1。8 本课题拟研究内容 6

2。 实验材料及方法 7

2。1 材料 7

2。1。1 菌种 7

2。1。2 质粒