Making the collection genome of Mycobacterium tuberculosis that are resistantto streptomycin in pUC19 vector: A model of search for genes responsible forresistance properties

[摘要] Tuberculosis caused by Mycobacterium tuberculosis (Mtb), which gives the infection is chronic and systematic, by spreading the human body through the lymphatic, vascular, respiratory, so it can attack any organ or tissue of the body. The purpose of this study to conduct a study on the gene responsible for resistance to antibiotics in bacteria Mtb that can be developed methods of detection of bacteria. This initial phase was the production of a collection of clones of E. coli bacteria that bring partial DNA (genome) Mtb bacteria that are resistant to the antibiotic streptomycin (strR) (partial genomic library). It is expected that the collection of the genome can be used as a model search for genes responsible for resistance to antibiotics, especially streptomycin. This study has been successfully created collection Mtb genome of bacteria that are resistant to streptomycin in the vector pUC19. The number of colonies that grew from a transformed bacterium E. coli JM101 was about 7000 colonies. Comparison of the number of colonies of white and blue is 1:4. Plasmid DNA isolation method based on alkaline lysis showed that of 41 recombinant plasmid isolated from white colonies, only 33 are visible containing DNA inserts. The data provides results ligation efficiency of 16.10%. The success of the manufacture of recombinant DNA is influenced by many things including comparisons between DNA ligation of vector and insert DNA, dephosphorylation of the ends of the DNA fragments to be ligated to avoid self-ligated. While the collection is influenced by its competent genome of the host cell used. The more competent host cell will provide much better results.

[学科分类] 药理学