260
FRIEDHELM HILDEBRANDT
Alternatively, Xenopus laevis oocytes may be microinjected with pools of
mRNA and assayed for expression of the desired protein. Pools of mRNA
are then divided into smaller and successively smaller fractions until a single clone is identified, that gives rise to the assayed function.
Positional cloning, differential cloning, etc.
If there is no prior knowledge available concerning protein sequence or
function, positional cloning (see Chap. 22) can be employed. In positional
cloning, cytogenetic abnormalities or linkage analysis point to a small genomic region in which the desired gene resides. This region is then defined
more narrowly by physical mapping techniques, and mutations in the responsible gene are identified by sequencing.
If celllines are available that express functional differences of interest,
differential cloning by subtractive hybridization or differential display can
be used.
References
Pesoie G, Attimonelli M, Liuni S (1988) A backtranslation method based on codon usage
strategy. Nucl Acids Res 16:1715-28
FRIEDHELM HILDEBRANDT
Alternatively, Xenopus laevis oocytes may be microinjected with pools of
mRNA and assayed for expression of the desired protein. Pools of mRNA
are then divided into smaller and successively smaller fractions until a single clone is identified, that gives rise to the assayed function.
Positional cloning, differential cloning, etc.
If there is no prior knowledge available concerning protein sequence or
function, positional cloning (see Chap. 22) can be employed. In positional
cloning, cytogenetic abnormalities or linkage analysis point to a small genomic region in which the desired gene resides. This region is then defined
more narrowly by physical mapping techniques, and mutations in the responsible gene are identified by sequencing.
If celllines are available that express functional differences of interest,
differential cloning by subtractive hybridization or differential display can
be used.
References
Pesoie G, Attimonelli M, Liuni S (1988) A backtranslation method based on codon usage
strategy. Nucl Acids Res 16:1715-28