植物内生细菌xg32菌株16srrna寡核苷酸探针杂交条件优化和特异.pdfVIP

34 1 () V o.l 34 N o. 1 20 11 3 JOURNAL OF NAN J ING NORMA L UN IVER SITY ( N a tura l Sc ence Ed t on) M ar, 2011 植物内生细菌XG 32菌株 16S rRNA 寡核苷酸 探针杂交条件优化和特异性验证 刘 佳, 王梅霞, 闫淑珍, 陈双林 ( , 2 10046) [] XG 32 , XG32 16S rRNA G enbank, BLA STDNA club O lgo , , 12 . , XG32: 35% , N aC l 12 mo l/ L, 42 , N aC l 002m ol/ L; XG32 , . , XG 32 . [] , , [ ] Q 9381 [] A [ ] 100 1-4 616( 20 11) 01-0089-07 Optim ization ofHybridization Conditions and Specific Verification for 16S rRNA O ligonucleotide Probe of Endop ytic Bacteria Strain XG32 L iu J ia, W ang M e ix ia, Y an S huzhen, Chen Shuang lin ( J angsu Eng n eer ng and T echnology R esearch Cen ter forM crob o logy, School of L fe Sc ences, N anj ng N orm al U n vers ty, N anj ng 2 10046, C h na) Abstract: F luorescence n s tu hybr d zat on w as appl ed to nvest ga te the clon ng and d str but on of the endophy t c bac- ter a stra n XG32 w th the ro le of grow th-prom ot ng and d sease contro l n plan ts. F rstly, a spec f c probe w as des gned theo re t ca lly at the base on the m d- spec f c sequence o f the 16S rRNA of the stra n XG 32 w th the G enBank da tabase, BLAST prog ram, DNA c lub so ftw a re and O l go softw are. Second ly, 12 stra ns w h ch sm lar o r adjacen t to the stra n XG32 w ere u sed to ce rt fy the spec f c ty of the probe by repeated hybr d zat on n the bas s o f the su tab le cond t ons o f hybr d zat on. T he results nd ca ted that appropr ate cond t ons fo r hybr d zat on o f the probe w ere 35% fo rmam de and 12 mo l/ L N aC l n hybr d za t on bu ffer, 42 hybr d za t on tem pera ture