猪带绦虫重组双歧杆菌疫苗不同保存条件下的稳定性研究

Stability of recombinant Bifidobacterium vaccine of Taenia solium under different preservation conditions

目的研究猪带绦虫TSO45W-4B基因、TSOL18基因和TSO45W-4B-TSOL18融合基因的重组双歧杆菌(Bb)疫苗在不同保存条件下的稳定性。方法分别将猪带绦虫重组质粒pGEX-TSO45W-4B、pGEX-TSOL18和pGEX-TSO45W-4BTSOL18电转入长双歧杆菌(B.longum),获得阳性菌株pGEX-TSO45W-4B/B.longum、pGEX-TSOL18/B.longum和pGEXTSO45W-4B-TSOL18/B.longum,分别置于不同条件下保存,取出菌液,抽提质粒,电泳检测,进行稳定性分析。结果阳性菌株pGEX-TSO45W-4B/B.longum、pGEX-TSOL18/B.longum和pGEX-TSO45W-4B-TSOL18/B.longum分别在常温下、4℃、-20℃、-80℃、-196℃(液氮中)分别保存48h、1周、1月、2月,抽提质粒,电泳发现阳性菌株pGEX-TSO45W-4B/B.longum、pGEX-TSOL18/B.longum和pGEX-TSO45W-4B-TSOL18/B.longum在液氮中、-80℃、-20℃中比较稳定,而在普通的室温环境下,很快发生质粒丢失现象。结论猪带绦虫TSO45W-4B基因、TSOL18基因和TSO45W-4B-TSOL18融合基因的重组Bb疫苗低温保存最稳定,常温最差,这为猪带绦虫重组Bb疫苗的进一步研究奠定了基础。

We studied the stability of the recombinant Bifidobacterium （Bb） vaccine of Taenia solium TSO45W-4B gene, TSOL18 gene or TSO45W-4B-TSOL18 fusion gene under different preservation conditions. The recombinant plasmid pGEX- TSO45W-4B, pGEX-TSOL18 or pGEX-TSO45W-4B-TSOL18 of Taenia solium was transformed into Bifidobacteriurn longurn （B. longum） to obtain positive strains pGEX-TSO45W-4B/B, longum, pGEX-TSOL18/B, longum or pGEX-TSO45W-4B- TSOL18/B. longum. At different preservation conditions, the stability of the positive strains was confirmed by electrophoresis of the extracted recombinant plasmid. The positive strains were preserved and the stability of the positive strains was con- firmed. With the condition of preservation at room temperature （RT）, 4 ℃, -20 ℃, -80 ℃, -196 ℃ （in liquid nitrogen） and for 48 hrs, one week, one month, two months, respectively, the plasmid electrophoresis demonstrated that the positive strains were more stable at -196 ℃, -80 ℃, and -20 ℃ while the plasmid disappeared at RT. It＇s indicated that the recombinant Bb vaccine of Taenia solium TSO45W-4B gene, TSOL18 gene or TSO45W 4B-TSOL18 fusion gene were more stable at lower pre- served temperature, which laid the foundation for the further study of the recombinant Bb vaccine of Taenia solium.