雞堆型艾美爾球蟲(chóng)乳酸脫氫酶增強(qiáng)型核酸疫苗構(gòu)建和免疫保護(hù)研究.pdf

1、ClassifiedIndex：Confidential(yes／no)：NoCode：10224NO1106355DissertationfortheMasterDegreeConstructionofImmune—enhancingDNAVaccineofEimeriaacervulinaLactateDehydrogenaseandItsImmuneProtectionCandidate：CongPeijunSupervisor：

2、ProfLiGuangxingDegreeCategory：MasterofAgricultureCollege：CollegeofVeterinaryMedicineFirstleveldiscipline：VeterinaryMedicineSecondleveldiscipline：BasicVeterinaryMedicineHarbinJuneChina2014東北農(nóng)業(yè)大學(xué)農(nóng)學(xué)碩士學(xué)位論文221堆型艾美爾球蟲(chóng)LDH基因克隆、測(cè)

3、序及序列比對(duì)16222堆型艾美爾球蟲(chóng)LDH基因的原核蛋白表達(dá)和純化20223LDH的多克隆抗體制備和生物活性檢測(cè)22224真核表達(dá)載體的構(gòu)建24225PVAX—IL181inkerLDH真核表達(dá)載體的構(gòu)建。25226重組質(zhì)粒體外瞬時(shí)表達(dá)的檢測(cè)一27227重組質(zhì)粒在雛雞體內(nèi)免疫效果檢測(cè)29228重組質(zhì)粒在對(duì)球蟲(chóng)感染的保護(hù)效果3023數(shù)據(jù)處理323結(jié)果3331RT—PCR擴(kuò)增EacervulinaLDH基因33311孢子化卵囊總RNA的提取

4、33312堆型艾美爾球蟲(chóng)LDH基因的擴(kuò)增33313重組克隆質(zhì)粒pMDl8TLDH鑒定34314重組克隆質(zhì)粒pMDl8TLDH的測(cè)序結(jié)果3432堆型艾美耳球蟲(chóng)LDH基因的原核表達(dá)及純化。35321LDH基因的亞克隆35322原核表達(dá)質(zhì)粒pET30aLDH的PCR鑒定35323重組質(zhì)粒pET30aLDH序列測(cè)定及二級(jí)結(jié)構(gòu)預(yù)測(cè)分析36324LDH基因原核蛋白表達(dá)及純化3733重組質(zhì)粒PVAXLDH的構(gòu)建和鑒定3934PVAXIL18LINKE

5、RLDH融合基因真核表達(dá)質(zhì)粒的構(gòu)建和鑒定39341PVAXIL181inker—LDH基因推測(cè)表達(dá)蛋白特性軟件預(yù)測(cè)結(jié)果39342重組質(zhì)粒PVAXIL181inkerLDH的PCR鑒定和酶切鑒定40343重組質(zhì)粒的測(cè)序鑒定4035重組質(zhì)粒的體外表達(dá)41351重組質(zhì)粒PVAXLDH體外瞬時(shí)表達(dá)41352重組質(zhì)粒PVAXIL181inkerLDH體外瞬時(shí)表達(dá)4136雛雞外周血T、B淋巴細(xì)胞增殖功能動(dòng)態(tài)變化42361雛雞外周血液T淋巴細(xì)胞增殖功