37, No. A transformant is a cell that has taken up additional DNA, usually a plasmid that confers some kind of antibiotic resistance. Routinely a first PCR screen of the transformants is based on the detection of stable integration of the selection marker gene. The selection for plasmid transformants utilizes their ampicillin resistance whereas selection for recombinant plasmids is based on the inactivation of the rpsL gene contained on the plasmid. Transformant cell is that cell in which transformation has occured. Still, in one experimental step, the recombinants among a number of randomly selected transformants can be selected and grown, by inoculation of small liquid cultures in two 24-wells plates. Figure 3: Blue-white color selection of recombinant bacteria using X-gal. Selection of recombinants due to inactivation of antibiotics is a laborious process as it requires:-(i) a vector with two antibiotic resistance marker (ii) preparation of two kinds of media plate, with one antibiotic each. Recombinant DNA Technology / By top. … The lacZ gene in the vector may sometimes be non-functional and may not produce β-galactosidase. Both (ampicillin medium containing) plates are inoculated with the same transformants, but one of the two plates contains additionally a second antibiotic: tetracyclin. Limitations of blue-white screening . A vector is chosen where … Selection and Screening of Transformants. Manjula R (2004) Positive selection system for identification of recombinants using α- complementation plasmids BioTechniques ,Vol. Selection of the transformed cells from the non-transformed population is done by using selectable marker genes. 6, pp. Selection methods in bacilli for recombinants and transformants of intra- and interspecific fused protoplasts. Analysis of transformants 1 Overview After the regeneration of protoplasts and two rounds of selection on antibiotic-containing medium the transformants can be analyzed for stable integration of the transgene by PCR-based methods. TRANSFORMATION: Process of introduction of cloning vector into the host cell. The blue-white technique is only a screening procedure; it is not a selection technique. Select the recombinant cells in the white colonies to culture. Selection by Antibiotic Resistance Genes. Akamatsu T, Sekiguchi J.


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