Expression of Chlamydophila psittaci MOMP heat-labile toxin B subunit fusion gene in transgenic rice.
A DNA fragment encoding the MOMP gene of Chlamydophila psittaci was fused to the heat-labile toxin B subunit gene (LTB-MOMP) and transferred into rice callus by Agrobacterium tumefaciens-mediated transformation. The LTB-MOMP fusion gene was detected in genomic DNA from transformed rice leaves by Southern blot and RT-PCR amplification. Synthesis and assembly of the LTB-MOMP fusion protein into pentamers was detected in transformed leaf extracts by immunoblot analysis. Binding of the pentamers to intestinal epithelial cell membrane glycolipid receptors was quantified by GM1-ganglioside enzyme-linked immunosorbent assay (GM1-ELISA). The ELISA results indicated that LTB-MOMP fusion protein made up 0.0033-0.0054% of the total soluble leaf protein. Meanwhile, this suggested that the fusion protein retained both its native antigenicity and the ability to form pentamers.
Zhang
Xiuxiang
X
Laboratory of Molecular Biology, College of Plant Science, Agricultural Division, Jilin University, 1068 Qinglong Road, Changchun 130062, PR China.
Yuan
Ziguo
Z
Guo
Xuejun
X
Li
Jingwen
J
Li
Zhaonan
Z
Wang
Qingyu
Q
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D023501:Chlamydophila / Q000235:genetics / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006358:Hot Temperature; D012275:Oryza; D010936:Plant Extracts / Q000235:genetics / Q000378:metabolism; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D010936:Plant Extracts; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins