根癌农杆菌介导甲型肝炎病毒(HAV)衣壳蛋白融合基因转化柑桔成年材料的研究

Agrobacterium-mediated Transformation of Citrus Mature Explants with Hepatitis a Virus Capsid Protein Fusion Gene

本研究以柑桔优良品种不知火(shinanui)为试料,通过携带质粒pBI121-A的根癌农杆菌转化成年树腋芽,获得转甲型肝炎病毒(HAV)衣壳蛋白融合基因不知火植株。研究结果表明,锦橙试管砧木培养15d时不知火成年树腋芽的嫁接成活率最高,达到90%;不知火不同成熟度的枝条应采用不同的消毒时间,木质化的枝条消毒时间要长些,半木质化的枝条消毒时间要短些。对获得的拟转化植株进行PCR分析以及对PCR阳性植株进行测序,结果表明HAV衣壳蛋白融合基因已整合到不知火的基因组中。

With the mature axillary buds of citrus excellent variety Shinanui （Citrus ReticulatexCitrus sinensis） severed as the explant, The plantlets which carried the hepatitis a virus capsid protein fusion gene were obtained through A grobacterium-mediated transformation. In this study, survival in graft of mature axillary buds of shinanui was the highest, up to 90%, when the orange rootstocks were cultured 15 days. Sterilization time was effected on maturity of shinanui branches. The more maturity of branches, the longer of sterilization time was. The putative transgenic plantlets was selected by PCR, PCR amplification product of PCR positive plantlets were sequenced, and the results showed that Hepatitis a virus capsid protein fusion gene was integrated into genome of transgenic shinanui.