The IPTG inducible expression vector containing the BYDV GAV coat protein gene was constructed and transferred into E.coli BL21(DE3).High level expression of the specific protein was achieved by IPTG induction.The results of SDS PAGE and Western blottong show that the expression product which accumulates 19.5% of the total cellular proteins estimated by scanning is 24 kD BYDV GAV coat protein plus eleven amino acids of pET 5a.
The Barley yellow dwarf virus (BYDV) GAV isolate was preserved at the Institute of Plant Protection of the Chinese Academy of Agricultural Sciences. The cDNA of BYDV GAV coat protein (CP) gene was amplified from the extracted RNA of BYDV GAV by using the polymerase chain reaction (PCR), and cloned into pGEM-7zf(+). Its complete nucleotide sequence has been determined by means of Sanger’s dideoxy-mediated chain-termination method. The result showed that BYDV GAV CP gene has 600nt. It shares 97.5% and 96.5% identity with CP gene of BYDV MAV-PS1 in terms of nucleotide and amino acid sequences respectively.
The universal primer of Luteovirus was designed and synthesized.An experimental system of RT PCR RFLP was developed in barley yellow dwarf virus (BYDVs).BYDVs can be distinguished qualitatively by RT PCR method.It was seen that different serotypes of BYDVs have critical different RFLP patters when the PCR producs were digested by restriction enzyme HinfI.The RFLP patterns of 7 isolates of PAV serotype were greatly different.These results indicate there existed sequence variations among different serotypes of BYDVs and vector phenotypes of PAV serotype.There is no difference between MAV serotypes in RFLP analysis.The slight distinction in the segment of coat protein gene of BYDVs can be revealed by RT PCR RFLP system.
以生物学测定和酶联检测鉴定出的大麦黄矮病毒 GAV 株系为材料,研究了病毒在麦二叉蚜体内的运行途径。在麦二叉蚜不同组织的超薄切片中,只有在唾液附腺组织中观察到膜包被的病毒粒体,在后肠腔和血淋巴中的病毒粒体是游离的。说明病毒与唾液附腺膜上传毒蛋白(受体)的相互识别是介导传播的根本原因,而麦蚜的获毒过程不存在专化的识别过程。
