[1]刘佳,王梅霞,闫淑珍,等.植物内生细菌XG32菌株16S rRNA寡核苷酸探针杂交条件优化和特异性验证[J].南京师大学报(自然科学版),2011,34(01):89-95.
　Liu Jia,Wang Meixia,Yan Shuzhen,et al.Optimization of Hybridization Conditions and Specific Verification for 16S rRNA Oligonucleotide Probe of Endophytic Bacteria Strain XG32[J].Journal of Nanjing Normal University(Natural Science Edition),2011,34(01):89-95.
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植物内生细菌XG32菌株16S rRNA寡核苷酸探针杂交条件优化和特异性验证()
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《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第34卷
期数:
2011年01期
页码:
89-95
栏目:
生命科学
出版日期:
2011-03-20

文章信息/Info

Title:
Optimization of Hybridization Conditions and Specific Verification for 16S rRNA Oligonucleotide Probe of Endophytic Bacteria Strain XG32
作者:
刘佳;王梅霞;闫淑珍;陈双林;
南京师范大学生命科学学院江苏省微生物工程技术研究中心, 江苏南京210046
Author(s):
Liu Jia; Wang Meixia; Yan Shuzhen; Chen Shuanglin
Jiangsu Engineering and Technology Research Center for Microbiology,School of Life Sciences,Nanjing Normal University,Nanjing 210046,China
关键词:
荧光原位杂交;  杂交条件优化;  特异探针验证
Keywords:
fluorescence in situ hybridization;  optim ization o f the hybr id ization conditions;  verification of spec ific probe
分类号:
Q938.1
摘要:
为了能利用荧光原位杂交方法来研究植物内生细菌XG32菌株在植物体内的定殖和分布,利用XG32菌株的16S rRNA中段特异性序列和Genbank数据库,用BLAST、DNAclub和Oligo软件设计了一个在理论上具有特异性的探针,在优化该探针的杂交条件基础上,通过与12个亲缘关系相邻和相近的菌株反复杂交来验证探针的特异性.结果显示,XG32探针适宜的杂交条件为:杂交缓冲液中甲酰胺浓度为35%,NaCl浓度为1.2 mol/L,杂交温度为42℃,清洗缓冲液中NaCl浓度为0.02 mol/L;探针与XG3探针.
Abstract:
F luorescence in situ hybr id ization w as applied to investiga te the cloning and distr ibution of the endophy tic bacter ia stra in XG32 w ith the ro le of grow th-prom oting and d isease contro l in plan ts. Firstly, a spec ific probe w as designed theo re tica lly at the base on the m id- spec ific sequence o f the 16S rRNA of the stra in XG32 w ith the GenBank da tabase, BLAST prog ram, DNAc lub so ftwa re and Oligo softw are. Second ly, 12 stra ins wh ich sim ilar o r adjacen t to the stra in XG32 w ere used to ce rtify the spec ific ity of the probe by repeated hybrid ization in the basis o f the su itab le cond itions o f hybridization. The results ind ica ted that appropriate cond itions fo r hybridization o f the probe w ere 35% fo rmam ide and 1.2 mo l/L NaC l in hybr idiza tion bu ffer, 42℃ hybr id iza tion tem pera ture, 0.02 mol/L NaC l in w ash ing buffer. The hyb rid ization signal was clear betw een the probe XG32 and the stra in XG32. To sum up, the probe XG32 w as the specific probe o f the strain XG32.

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备注/Memo

备注/Memo:
基金项目: 江苏省高校自然科学基础研究项目( 07K JB180063 ). 通讯联系人: 闫淑珍, 副教授, 研究方向: 微生物的应用. E-m ail:yanshuzhen@njnu. edu. cn
更新日期/Last Update: 2013-04-11