Transformation of Agrobacterium Tumefactions by Conjugation Technique

Authors : Raghad N G ALZAIDY, Najwa I ALBARHAWI, Abdullah N ALNIEMI

Pages : 28-39

View : 59 | Download : 11

Publication Date : 2018-12-01

Article Type : Research Paper

Abstract :Two genera of Rhizobiaceae ,wild strain Agrobacterium tumefaciens insert ignore into journalissuearticles values(AtMcol);  and standerd strain Agrobacterium tumefaciens insert ignore into journalissuearticles values(C58C1);, in addition to Sinrhizobium melilti are used in this study. The results showed, these two strains ofA. tumifaciens bacteria were resistant to Trimetheprim and sensitive to chloromphinicol while the S.melilti bacteria gave the opposite of these results,and used as genetic markers for conjugation as one of the methods used in the technique of Horizontal gene transfer, with the use of acardine orange to cure the bacteria from plasmid.Two conjugation procces were performed, between Agrobacterium tumefaciens insert ignore into journalissuearticles values(AtMcol); and S. melilti, and between Agrobacterium tumefaciens insert ignore into journalissuearticles values(C58C1);and S.melilti, result showed succecful of conjugation procces with conjugation frequency 1.8x10-2 and 2.1x10-2 respectively, and the transconjugant bacteria result from procces succeded in infecting the root tip of alfalfa seedlings to form root nodules. Characterization of plasmid DNA content for bacterial  isolates belonging to conjugated cells, were carried out by using agarose gel electrophorsis and the results showed isolate the presence of discrete clear and isolated band of similar size present in agaros gel.
Keywords : Conjugation technique, Biochemical, DNA

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