微小隱孢子蟲Cp23基因在短乳桿菌S-層蛋白中的嵌合型原核表達載體的構建.pdf

1、 分類號 Q75 學校代碼 10129 U D C 577 學 號 2009210028 微小隱孢子蟲 Cp23 基因在短乳桿菌 S-層 蛋白中的嵌合型原核表達載體的構建 Construction of Chimeric Prokaryotic Expression Vector of Cp23 G

2、ene from Cryptosporidium parvum into slp Gene from Lactobacillus brevis 申 請 人：蘭 麗 學科門類：理 學 學科專業(yè)：生物化學與分子生物學 研究方向：蛋白質化學 指導教師：魏 建 民 教授 格日勒圖 教授 論文提交日期：二〇一二年五月 Construction of Chimeric Prokaryotic Expression Vector of Cp

3、23 Gene from Cryptosporidium parvum into slp Gene from Lactobacillus brevis Abstract S-layer protein is usually located in the outer surface of cell wall or cell membrane of bacteria and archaea with single-molecule and

4、Lattice-like structure . These proteins has a unique inherent properties which easily ruptured by chemicals, than protein crystals can be reassembled into the original lattice. Its function is maintain the shape of the b

5、acterial cells, combined with the high molecular enzymes, adhesion and invasion the target cells. It is an important structure for the bacteria to adapt to the environment. Some Lactobacillus S-layer protein with molecul

6、ar weight between 25-71KDa is currently the smallest known S-layer protein. Cryptosporidium parvum parasitic in humans and mammals which can cause cryptosporidiosis . The pathogenic and immune-related antigen protein exi

7、st in its every complex life cycle . 30 kinds of promising vaccine candidate molecules have been reported, such as GP900, CSL, Cp23, Cp15/40/60, Cp12 and Cp21.Because of the conservative and stronger immune activity, the

8、 Cryptosporidium sporozoite surface antigen which encoded by Cp23 gene sequence is considered as the best target antigen of Cryptosporidium parvum in immunodiagnosis and vaccines developed by most scholars. This study us

9、es molecular biology techniques to construct the chimeric expression vector with surface antigen Cp23 gene and the S-layer protein gene of Lactobacillus brevis. Lay the foundation for the achievement of the gene fusion j

10、oint development of the vaccine vector. The experimental results are as follows: 1 Survey and identification the S-layer protein and slp gene of the lactic acid bacterias by SDS-PAGE and PCR respectively in protein and n

11、ucleic acid level. Detect the size of the S-layer protein of Lactobacillus brevis is approximately 47kDa. 2 Analyzed the slp gene sequences of Lactobacillus brevis through the tools such as Genetyx and Bioinf . The seque

12、ncing results showed that the effective nucleotide sequence of Lactobacillus brevis is 1263bp which deduced 421 amino acids, and determine the outergene into the exact sites. 3 The recombinant plasmid pET30a-slp-P23 was