转移猪轮状病毒VP4基因玉米植株的获得

根据GenBank中猪轮状病毒VP4基因序列设计引物，从重组克隆载体pMD18-T-VP4中PCR扩增VP4基因，将其插入到植物表达载体pCAMBIA3301中CaMV35S启动子下游，构建成高效植物表达载体pCAMBIA3301－VP4；然后利用农杆菌介导法，以玉米自交系H99茎尖来源的玉米愈伤组织为材料，将pCAMBIA3301－VP4导入其中，分化诱导获得再生苗后，对其进行PCR、Southern杂交和RT-PCR检测。结果表明，外源目的基因VP4已成功地转入玉米基因组中，并且目的基因能在转基因玉米植株中获得转录。

Porcine rotavirus VP4 gene transferred into maize

According to the sequence of VP4 in GenBank,a pair of primer for amplifying the VP4 gene has been de- signed,with which the predicted fragment had been amplified by PCR from pMD18-T-VP4 plasmid. The VP4 gene was cloned into pCAMBIA3301 plasmid, the recombinant plasmid pCAMBIA3301-VP4 was constructed. Then the correct recombinants were transformed into callus of maize inbred lines H99 induced by Agrobacterium tumefaciens EHA101. The expression of VP4 in transgenic maize were examined by PCR, Southern blotting detection and RT- PCR. The result showed that the exogenous VP4 gene had been integrated into maize genome via PCR analysis and had been expressed by RT-PCR analysis.