表达pedv保护性抗原靶向融合肽乳酸杆菌活菌载体的构建及其免疫原性分析兽医学预防兽医学专业论文.docxVIP

CONTENTS量量量曼曼曼曼曼曼皇量量昌舅詈量量量量量曼量量量置皇量量舅量量詈皇鲁昌昌置量鼍量罾量量曼量曼曼曼I CONTENTS 量量量曼曼曼曼曼曼皇量量昌舅詈量量量量量曼量量量置皇量量舅量量詈皇鲁昌昌置量鼍量罾量量曼量曼曼曼I I鼍I CoNTENTS Abstract in Chinese ． ． ． ．．． ． ．． ． ． ． ． ． ．I Abstract in English ． ． ． ．． ．． ． ． ．III 1 Introduction ．．．． ． ． ． ． ． ． ． ．． ． ． ．1 1．1 Prevalence and hazard of PED ．．． ． ． ．． ． ． ．． ． ．． ．．．1 1．2 Eptiology of PEDV ． ． ． ． ·· ·”2 1．2．1 Pathogen classification and main biological characteristics ．．．．． ·”2 1．2．2 Genome structure and eoding protein ． ． ．． ． -·· 一2 1．3 Pathogenicity and main pathological features of PEDV ．．．． - · ·4 1。4 Research progress of novel PEDV vaccine based S gene 4 1．5 Progress M cell targeting vaccine 4 1．6 Progress dendritic cell targeting vaccine ． · - 5 1．7 Progress Expression Foreign Antigen Gene by Lactic Acid Bacteria(LAB) ·6 1．8 Purpose and Significance of the Investigation 7 2 Materials and methods 8 2．1 Materials 8 2．1．1 Virus．strain and plasmid ·8 2．1．2 Experimental animal 8 2．1．3 Primer ．8 2．1．4 Reagent ·9 2．1．5 Main laboratory apparatus and devices ·y 2．2 Empirical method 9 2．2．1 Construction ofrecombinant L．casei experssing fusion gene ． ”9 2．2．2 Protein expression and identification ofrecombinant L．casei ．． ． · ·18 2．2．3 Construction，protein expression and purification of recombinant E．coli ”1 9 2．2．4 The immunity effective evaluation of dose immune by recombinant￡．casei ·21 2．2．6 The immunity effective evaluation of twice immune by recombinant L．casei ·23 3 ResuIts ．．25 3．1 The result of amplification of S 1 gene via PCR z， 3．2 The result of amplification of ps420’gene via PCR．． ．． 25 3．3 The result ofamplification oftarget gene via PCR ． ． ． - 26 3．4 Identification of recombinant L．casei ． 20 3．4．1 Identification ofrecombinant expression vector pPG-T7910-PPT-ps420 26 3．4．2 Identification ofrecombinant expression vector pPG·T7910一PPT-ps420-c01 一27 3．4．3 Identification ofrecombinant expression vector pPG—TTgl0-PPT-ps420·DCpep 28 3．4．4 Identification of recombinant expressio