应用改进的多克隆抗体Western blot技术研究柑桔速衰病毒蛋白(英文)  

DEVELOPMENT OF WESTERN BLOT PROCEDURE FOR USING POLYCLONAL ANTIBODIES TO STUDY THE PROTEINS OF CITRUS TRISTEZA VIRUS

作　　者：林尤剑[1] 谢联辉[1] PhyllisA.Rundell CharlesA.Powell 

机构地区：[1]福建农业大学植物病毒研究所,福州350002 [2]佛罗里达大学印第安那河研究和教育中心

出　　处：《植物病理学报》2000年第3期250-256,共7页Acta Phytopathologica Sinica

摘　　要：本文利用多克隆抗体发展了一种无背景的 Western blot技术并用于研究柑桔速衰病毒( CTV)的蛋白。结果表明 ,利用 CTV兔多克隆抗体 12 12和 10 52发展的 Western blot技术可以检测到感染 CTV的墨西哥酸橙或 Citrusexcelsa植株内 CTV的 4种蛋白 P1、P2、P3和 P4。在健株或病株内杂交反应均无非特异性背景。从不同寄主上分离到的 CTV不同分离物的蛋白条带是不同的。利用12 12和 10 52抗体均可以检测到感染 6个 CTV分离物的墨西哥酸橙幼苗内的 P1、P2和 P3。利用10 52抗体能检测到感染严重型分离物 T36、T3和 Mm2的墨西哥酸橙幼苗内微弱的 P4 ,但感染轻型分离物 T30、T2 6和 T4的幼苗内则检测不到。利用 12 12抗体检测不到 P4。在 C.excelsa内 ,12 12和10 52抗体均能检测到感染所有分离物的病株内的 P1。在感染 T3、T2 6、T4或 Mm2的病株内能检测到 P2 ,但在感染 T30和 36分离物的病株内则检测不到。在感染 T36、T3、T2 6、T4和 Mm2的病株内可检测到 P3,但在感染 T30的病株内则检测不到。在大多数植物内 ,P1、P2、P3和 P4的分子量分别约为 2 5k Da,2 4 k Da,2 1k Da和 18k Da。在感染 T36分离物的 C. excelsa植株体内 ,P1和 P3的分子量分别约为 2 7k Da和 2 2 k Da,比感染其它分离物的 C.excelsa和墨西哥酸?A non back ground reaction Western blot procedure for using polyclonal antibodies to study the proteins of citrus tristeza virus(CTV) was developed. The results showed four proteins, named P1, P2, P3 and P4, of CTV were detected in Mexican lime or Citrus excelsa plants that were infected with CTV by the Western blot procedure with CTV rabbit polyclonal antibodies, 1212 and 1052. No non specific back ground reactions were detected in the healthy or diseased plants. The patterns of specific proteins of different isolates of CTV in different hosts were different. The P1, P2 and P3 proteins were observed in Mexican lime seedlings infected with 6 isolates of CTV by 1212 and 1052. Antibody 1052 detected a weak P4 band in blots from Mexican lime seedlings infected with the severe isolates, T36, T3 and Mm2, but not the mild isolates, T30, T26 and T4. Antibody 1212 does not detect P4. In Citrus excelsa , both the 1212 and 1052 antibodies detected P1 in trees infected with all isolates. P2 was detec ted in trees infected with T3, T26, T4 or Mm2, but not in trees infected with T30 and T36. P3 was detected in trees infected with T36, T3, T26, T4 and Mm2, but not T30. The molecular weights of P1, P2, P3, and P4 of CTV in most plants were about 25 kDa, 24 kDa, 21 kDa and 18 kDa, respectively. The molecular weights of P1 and P3 of T36 in Citrus excelsa plants were about 27 kDa and 22 kDa, respectively, a little larger than that of other isolates in both the Mexican lime and Citrus excelsa plants. Molecular weights of the P1, P2 and P3 were the same as those of CP, CP1 and CP2 detected by monoclonal antibodies. Therefore, the P1, P2 and P3 are probably the CTV coat proteins, CP, CP1 and CP2, respectively. The nature of P4 is unknown. The results also indicated that the Western blot procedure is a useful tool for studying CTV with CTV specific polyclonal antibodies. With the Western blot procedure, different isolates or strains of CTV could be distinguished by analysis of the specific protein patterns of

关 键 词：WESTERN-BLOT 多克隆抗体 柑桔速衰病毒 蛋白 

分 类 号：S436.661.1[农业科学—农业昆虫与害虫防治]