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将生长抑素(SS)和乙肝表面抗原基因(HBsAg)融合后,插入真核表达载体pcDNA3,构建成pcDNA3-SS,再转化减毒沙门氏菌(S.typhimurium,dam-和phop-),构建了以减毒沙门氏菌为载体的生长抑素口服DNA疫苗ZJ111/pcDNA3-SS,并通过体外传代、灌服草鱼检验了ZJ111/pcDNA3-SS的侵袭力及体外和侵入草鱼体内过程中的稳定性。对草鱼肝脏和脾脏分离菌的生化特性检验和特异性PCR鉴定表明,ZJ111/pcDNA3-SS能很好地侵入草鱼体内;对在Amp+、Amp-平板上传5、10代和灌服7d后草鱼肝、脾脏分离的减毒菌抽提重组质粒进行PCR和酶切鉴定表明,减毒菌中的重组质粒在体外和侵入草鱼体内过程中具有较好的稳定性。
After fusion of somatostatin (SS) and hepatitis B surface antigen gene (HBsAg), the eukaryotic expression vector pcDNA3 was inserted into pcDNA3-SS and transformed into S. typhimurium (dam- and phop-) The attenuated Salmonella-mediated somatostatin oral DNA vaccine ZJ111 / pcDNA3-SS was used to test the invasiveness of ZJ111 / pcDNA3-SS and the stability in in vivo and in vitro invasion of grass carp. The biochemical characterization and specific PCR analysis of the liver and spleen isolates from grass carp showed that ZJ111 / pcDNA3-SS could invade well into the grass carp. The expression of ZJ111 / pcDNA3- , The attenuated spleen isolates were extracted by recombinant plasmid PCR and restriction enzyme digestion showed that the attenuated bacteria in recombinant plasmids in vitro and invade grass carp in the process has good stability.