Construction and characterization of a papaya BAC library as a foundation for molecular dissection of a tree-fruit genome |
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Authors: | R Ming P H Moore F Zee C A Abbey H Ma A H Paterson |
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Institution: | (1) Hawaii Agriculture Research Center, Aiea, HI 96701, USA e-mail: rming@harc-hspa.com, US;(2) USDA-ARS, Tropical Fruit, Vegetable and Sugarcane Research Unit, Aiea, HI 96701, USA, US;(3) USDA-ARS, National Clonal Germplasm Repository, Hilo, HI 96720, USA, US;(4) Dept Soil and Crop Science, Texas A&M University, College Station, TX 77843, USA Present address: A.H. Paterson, Applied Genetic Technology Center, University of Georgia, Athens, GA 30602, USA, US;(5) Dept Molecular Biosciences and Biosystems Engineering, University of Hawaii, Honolulu, HI 96822, USA, US |
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Abstract: | A bacterial artificial chromosome (BAC) library was constructed from high-molecular-weight DNA isolated from young leaves
of papaya (Carica papaya L.). This BAC library consists of 39168 clones from two separate ligation reactions. The average insert size of the library
is 132 kb; 96.5% of the 18700 clones from the first ligation contained inserts that averaged 86 kb in size, 95.7% of the 20468
clones from the second ligation contained inserts that averaged 174 kb in size. Two sorghum chloroplast probes hybridized
separately to the library and revealed a total of 504 chloroplast clones or 1.4% of the library. The entire BAC library was
estimated to provide 13.7× papaya-genome equivalents, excluding the false-positive and chloroplast clones. High-density filters
were made containing 94% or 36864 clones of the library with 12.7× papaya-genome equivalents. Eleven papaya-cDNA and ten Arabidopsis-cDNA probes detected an average of 22.8 BACs per probe in the library. Because of its relatively small genome (372 Mbp/1
C) and its ability to produce ripe fruit 9 to 15 months after planting, papaya shows promise as a model plant for studying
genes that affect fruiting characters. A rapid approach to locating fruit-controlling genes will be to assemble a physical
map based on BAC contigs to which ESTs have hybridized. A physical map of the papaya genome will significantly enhance our
capacity to clone and manipulate genes of economic importance.
Received: 11 April 2000 / Accepted: 28 July 2000 |
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Keywords: | Carica Genome Physical mapping Positional cloning |
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