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[1]莫才清,周俊初,李阜棣.含发光酶基因的转座子质粒载体的改造及向根瘤菌HN01的转座[J].应用与环境生物学报,1997,3(03):252-257.
 Mo Caiqing,Zhou Junchu,Li Fudi.DEVELOPMENT OF A SUICIDE PLASMID CONTAINING luxAB GENES AND ITS TRANSLOCATION TO RHIZOBIUM FREDII HN01[J].Chinese Journal of Applied & Environmental Biology,1997,3(03):252-257.
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含发光酶基因的转座子质粒载体的改造及向根瘤菌HN01的转座()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
3卷
期数:
1997年03期
页码:
252-257
栏目:
论文
出版日期:
1997-08-25

文章信息/Info

Title:
DEVELOPMENT OF A SUICIDE PLASMID CONTAINING luxAB GENES AND ITS TRANSLOCATION TO RHIZOBIUM FREDII HN01
作者:
莫才清 周俊初 李阜棣
华中农业大学生命科学院 武汉 430070
Author(s):
Mo Caiqing Zhou Junchu Li Fudi
Life-Science College, Huazhong Agri Univ, Wuhan 430070
关键词:
发光酶基因大豆根瘤菌标记根瘤菌
Keywords:
luxAB geneRhizobium frediimark rhizobia
摘要:
将从自杀性重组质粒pDB30上分离的3.6kb含卡那霉素抗性基因和发光酶基因(luxAB)的BglⅡ酶切片段,克隆到小Mr的高拷贝质粒pIJ2925上,构成新的质粒,pHNLX1011;然后将pHNLX1011经BglⅡ部分酶切,酶连在含蔗糖酶基因(sacB)的自杀性重组质粒载体pMH1701上,载体分别采用BglⅡ酶切和BamHⅠ酶切.连接转化后共得到4种重组质粒类型的菌株,它们具有不同发光活性;将它们分别与快生型大豆根瘤菌HN01进行三亲本杂交,发现它们的转座频率均较pDB30高.对经转座标记后的根瘤菌进行发光活性检测,只有pHNC3质粒所转座的根瘤菌浇具有较强发光活性.所得发光根瘤菌衍生菌株能在野大豆和栽培大豆黑农39根际形成正常根瘤,能用X光片记录下根瘤的发光活性.
Abstract:
A 3.6 kb fragment containing Km resistance and luxAB genes was isolated from a suicide plasmid pDBO by Bgl Ⅱ digestion,then cloned into a high copy number plasmid plJ2925 and formed pHNLX1011. The plasmid was digested and inserted into a new suicide vector pMH1701, which was digested by Bgl Ⅱor BamH Ⅱ, respectively. The transformants included four kinds of plasmid type with different luminescence activity. They were conjugated with Rhizobium fredii AN01 at the help of pRK2013. It was found that the four plasmids had higher translocation frequency than that of pDB30.The luminescence activity of the HN01 conjugates was tested and indicated that only the colonies formed by pHNC3 had high lumines-cence activity. The Rlizobum fredii HN01 luminescence derivaitives could form normal nodules on the root of wild soybeans and cultivar soybean Heinong 39,and the luminescence activity of nodules could be recorded by X-ray film.

参考文献/References:

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备注/Memo

备注/Memo:
收稿日期:1996-3-11;接受日期:1996-7-16。
基金项目:国家“863”项目,国家自然科学基金。
更新日期/Last Update: 1900-01-01