To express a cloned gene (for a desired product) in a transformed cell means that it has to transcript and translate. Sometimes functional proteins are produced by post-translation modifications. Protein synthesis is based on the encoded information in the sequence of the DNA strand bases. Structural gene directs the cells machinery for sequential linear arrangements
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The HRT method is utilised for recombinants without introns. The most difficult step in applying hybridisation is obtaining a suitable probe. If the target DNA is represented with highly abundant mRNA, a small number of cDNA clones can be screened and the target gene can be identified by sequencing or hybrid released translation. In HRT,
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The replica plate method is used in genetic engineering for identifying transformed bacterial cells (cells containing foreign DNA). Only some of the cells transformed with a specific DNA may actually have foreign DNA. Thus, it is essential to screen those cells having foreign DNA by the technique of colony hybridisation (only for bacterial cells) figure.
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For conventional DNA sequencing templates of signal stranded DNAs are used. PCR-based methods can be employed to manufacture them. Also vectors based on certain bacteriophages whose genomes assume a single stranded DNA at some stages in their life cycle can be used. M13 Bacteriophage M13 is a filamentous bacteriophage. Its genomes bear single-stranded circular DNA
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