PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 91pg1

IDENTIFYING NOVEL MAIZE GENES BY PARTIAL SEQUENCING OF RANDOMLY SELECTED COMPLEMENTARY DNAS

Cindy Rixmann, Sharada Vijaychander, Anitha Chandrasekhar, Carlyn Keith, Danee Hoang, Medha Joshi, Leah Quintana, Pisanee Sopapan and Chris Baysdorfer, Department of Biological Sciences, California State University, Hayward, CA 94542


We are interested in functionally identifying novel maize genes. In this study we have partially (end) sequenced cDNAs randomly selected from a mature leaf library. This library contains a high proportion of unidirectionally cloned, mid to full length cDNAs with a low representation of abundant leaf mRNAs. A single PCR protocol is apable of amplifying most (>80%) cDNAs. At present we have sequenced 300+ cDNAs with the following results: previously sequenced maize cDNAs = 5%; cDNAs provisionally identified on the basis of sequence similarity to previously sequenced genes from organisms other than maize = 15%; unidentified cDNAs = 80%. At this point in the screening only 2% of the cDNAs are duplicates. Sequence and similarity data for the provisionally identified cDNAs will be presented. About one half of the provisionally identified cDNAs consist of representatives of photosynthetic metabolism, glycolysis, the citric acid cycle and protein synthesis. Approximately a fifth encode nucleic acid binding proteins. The remaining cDNAs encode proteins with widely diverse functions. Two thirds of the provisionally identified cDNAs either were, or could have been, identified entirely as a result of sequence similarity to non-plant genes. An analysis of GenBank/EMBI., submissions during the last two years shows that new submissions to the databases account for a small percentage of provisionally identified genes. We believe that the results of this and other cDNA sequencing projects show that this approach offers a rapid and inexpensive way to functionally identify large numbers of plant genes.


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