Differential display-mediated rapid identification of different members of a multigene family, HSP 16.9 in wheat.

Abstract:

:Isolation of cDNAs encoding individual members of a gene family is essential for assessing their role in a biological phenomenon. However, this process is often laborious and slow due to highly conserved protein-coding region that interferes with the isolation of the individual members. Identification of gene-specific probes from 3' non-coding regions of different members can assist in the fast retrieval and characterization of individual members of a multigene family. We used the recent technique of differential display for the same purpose. As an example of a multigene family in plants, we selected a heat shock protein gene family, HSP16.9 from wheat, with estimated 12 members. We modified the original differential display technique for selective amplification of the 3' non-coding regions of different wheat HSP16.9 genes by replacing the random 10-mer in the original method with a conserved HSP16.9 gene family-specific primer. Sixteen cDNA fragments from these experiments were sequenced and they represent 8 different members of a 12 member gene family. Our success can be attributed to shorter 3' non-coding regions that are typical of higher-plant genes and use of highly conserved gene family-specific primer in these experiments. This modified differential display technique can be of general application to other plant systems where cloning of the different members of a gene family is desired.

journal_name

Plant Mol Biol

journal_title

Plant molecular biology

authors

Joshi CP,Nguyen HT

doi

10.1007/BF00042230

subject

Has Abstract

pub_date

1996-06-01 00:00:00

pages

575-84

issue

3

eissn

0167-4412

issn

1573-5028

journal_volume

31

pub_type

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