Parental genes

Figure 2.5 Scheme for DNA shuffling illustrated for the case in which the parent genes originate from the WT by some form of mutagenesis [7e],... 

A modified version of this protocol, called THIO-ITCHY, includes the random incorporation of a-phosphothioate nucleotide analogs into the parent genes [27,28]. Exonuclease III activity is inhibited at sites of analog incorporation, which relieves the efforts of producing incremental truncation aliquots. In combination with epPCR, the diversity of the fusion libraries created can be further expanded. 

Fig. 3.2. Schematic representation of DNA shuffling using 2 parental genes.

Digest plasmids (5 pg) containing parent gene with appropriate restriction endonucleases. 

Electrophorese and recover DNA fragments of parent gene from agarose gels using gel extraction kit (see Section 3.2.1). 

The preparation of parent genes by PCR is effective when it is difficult to excise... 

Genomic imprinting is the mechanism by, which only one of the two parental gene copies either maternal or paternal—is expressed. Modifications in DNA and histones are the marks of genomic imprinting. There are about 50 known imprinted genes in the human genome (64). For example, the... 

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