[Cloning, expression of the CP gene and antiserum preparation of zucchini yellow mosaic virus infecting Benincasa hispida Cogn. var. chieh-qua How].
A Guangzhou isolate of ZYMV infecting Benincasa hispida Cogn. var. chieh-qua How was identified by indicator tests and partial sequence amplification. The coat protein (CP) gene of this virus was amplified by RT-PCR, and ligated to the expression vector pET-22b(+). The recombinant plasmid pET-ZCP was transformed into E. coli BL21 (DE3) and then induced to express by IPTG. It was shown that the CP gene was highly expressed by SDS-PAGE and Western blot analysis. The molecular weight of the recombinant protein was about 33.0 kD. Antiserum with high specificity was produced after the rabbit was immunized with purified recombinant protein, and the titer was determined to be 1/4096 by antigen coating plate-ELISA (ACP-ELISA).
Song
Li-min
LM
Department of Plant Pathology, China Agricultural University, Beijing 100094, China. lm-song@sohu.com
Lu
Cai-ge
CG
Liang
Wen-xing
WX
Huang
Jin-guang
JG
Li
Huai-fang
HF
D000818:Animals; D036022:Capsid Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D003001:Cloning, Molecular; D028464:Cucurbita / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D007106:Immune Sera; D017800:Potyvirus / Q000235:genetics* / Q000276:immunology; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction
D036022:Capsid Proteins; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; C086081:coat protein, Zucchini yellow mosaic virus