Construction of eukaryotic expression vector of outer membrane protein P30 from Mycoplasma oumvipneoniae and its transient expression in tobacco
In this study,the gene of the outer membrane protein P30 with the length of 798 bp was amplified from the genome of Mycoplasma oumvipneoniae(MO) Y-98 using a pair of specific primers and was cloned into the pMD19-T Simple vector.After the sequence was confirmed,the MOP30 gene was extracted by digesting the recombinant plasmid,and was subcloned into the pCAMBIA1300-YFP vector to construct eukaryotic expression vector named pCAMBIA1300-MOP30-YFP.Then the constructed plasmid pCAMBIA1300-MOP30-YFP was transformed Agrobacterium competent cells,and infected tobacco leaves.The enhanced yellow fluorescent protein(YFP) was observed under imaging microscope,and the mRNA of MOP30 was detected by RT-PCR.Western-blotting showed a 57 000 protein-specific band.The results indicated that the vector pCAMBIA1300-MOP30-YFP was successfully constructed and expressed in tobacco,laying a foundation of further investigation of transgenic plant vaccines for preventing Mycoplasma pneumonia of sheep.
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万方数据
维普
