畜牧兽医杂志
畜牧獸醫雜誌
축목수의잡지
JOURNAL OF ANIMAL SCIAENCE AND VETERINARY MEDICINE
2012年
4期
41-44
,共4页
猪圆环病毒2型%Cap%a—IFN%体液免疫%细胞免疫
豬圓環病毒2型%Cap%a—IFN%體液免疫%細胞免疫
저원배병독2형%Cap%a—IFN%체액면역%세포면역
Porcine Circovirus Type 2%Cap%α-IFN%h umoral immunity%cellumar immunity
利用PCR技术和亚克隆技术构建了猪圆环病毒2型（PCV2）Cap蛋白基因与猪a—IFN基因的真核双表达质粒pIRES—Cap—α—IFN，经酶切、PCR鉴定和测序表明构建成功。将该质粒免疫6～8周龄BALB／c纯系小鼠，每隔2周免疫1次，共免2次，每次免疫前采血分离血清，实验结束时杀鼠取脾制备T淋巴细胞。用间接ELISA方法检测小鼠血清抗体水平，用MTT法检测T细胞增殖情况。结果表明，pIRES-Cap—α—IFN免疫组能显著刺激T淋巴细胞增殖（P〈0．05），促进机体产生体液免疫但差异不显著（P〉0．05），为PCV2核酸疫苗研究奠定了较好的前期基础。
利用PCR技術和亞剋隆技術構建瞭豬圓環病毒2型（PCV2）Cap蛋白基因與豬a—IFN基因的真覈雙錶達質粒pIRES—Cap—α—IFN，經酶切、PCR鑒定和測序錶明構建成功。將該質粒免疫6～8週齡BALB／c純繫小鼠，每隔2週免疫1次，共免2次，每次免疫前採血分離血清，實驗結束時殺鼠取脾製備T淋巴細胞。用間接ELISA方法檢測小鼠血清抗體水平，用MTT法檢測T細胞增殖情況。結果錶明，pIRES-Cap—α—IFN免疫組能顯著刺激T淋巴細胞增殖（P〈0．05），促進機體產生體液免疫但差異不顯著（P〉0．05），為PCV2覈痠疫苗研究奠定瞭較好的前期基礎。
이용PCR기술화아극륭기술구건료저원배병독2형（PCV2）Cap단백기인여저a—IFN기인적진핵쌍표체질립pIRES—Cap—α—IFN，경매절、PCR감정화측서표명구건성공。장해질립면역6～8주령BALB／c순계소서，매격2주면역1차，공면2차，매차면역전채혈분리혈청，실험결속시살서취비제비T림파세포。용간접ELISA방법검측소서혈청항체수평，용MTT법검측T세포증식정황。결과표명，pIRES-Cap—α—IFN면역조능현저자격T림파세포증식（P〈0．05），촉진궤체산생체액면역단차이불현저（P〉0．05），위PCV2핵산역묘연구전정료교호적전기기출。
Recombinant plasmid pIRES-Cap-IFN-α was constructed using Cap gene of PCV2 and porcine IFN-α gene,and was used to immunize mice. The plasmid was used to immune 6-8 weeks age BALB/c mice, once time every two weeks. Serum was separated before immunity, T lymphocyte was prepared with mice spleen at the end of the experiment. T lymphocyte proliferation was examined with MTT method and antibody levels were detected using indirect ELISA. The results showed that the recombinant plasmid pIRES-Cap-IFN-α stimulated lymphocyte proliferation（P〈0.05）and induced the humoral immune response （P〈0.05）. It laid the foundation for further study and played a great role for the PCV2 subunit vaccine research.
