TY - JOUR
T1 - Construction and characterization of a common bean bacterial artificial chromosome library
AU - Vanhouten, Wim
AU - MacKenzie, Sally
N1 - Funding Information:
We thank Drs N.D. Young and D. Danesh for providing the vector pECSBAC4, and for helpful suggestions during construction of the library. We thank Drs R. Wing and D. Frisch for assistance in the preparation of high-density colony filters. This work was supported by grants from the USDA-NRI (64-37300-0451) and the NIH (R21 GM 54154-01).
PY - 1999/8
Y1 - 1999/8
N2 - We have constructed a common bean (Phaseolus vulgaris L.) bacterial artificial chromosome (BAC) library consisting of 33 792 clones and an estimated 3- to 5-fold coverage of the common bean genome. Leaf nuclei were used as the source for high-molecular-weight DNA, and an endonuclease/methylase competition assay was employed to partially cleave the DNA. The library was screened with a number of nuclear and mitochondrial probes. Each nuclear probe identified at least two BACs with an average insert size of ca. 100 kb. Only 26 clones were identified after hybridizing with mitochondrial probes, indicating contamination with organellar sequences is low. Numerous clones could be identified after screening the library with two repetitive probes flanking the nuclear fertility restorer Fr. Intriguingly, 12 clones appeared to hybridize to both markers, and restriction analysis of these clones revealed that they can be assembled into maximally four contigs, suggesting that these repetitive probes may be useful for the physical mapping of the Fr locus.
AB - We have constructed a common bean (Phaseolus vulgaris L.) bacterial artificial chromosome (BAC) library consisting of 33 792 clones and an estimated 3- to 5-fold coverage of the common bean genome. Leaf nuclei were used as the source for high-molecular-weight DNA, and an endonuclease/methylase competition assay was employed to partially cleave the DNA. The library was screened with a number of nuclear and mitochondrial probes. Each nuclear probe identified at least two BACs with an average insert size of ca. 100 kb. Only 26 clones were identified after hybridizing with mitochondrial probes, indicating contamination with organellar sequences is low. Numerous clones could be identified after screening the library with two repetitive probes flanking the nuclear fertility restorer Fr. Intriguingly, 12 clones appeared to hybridize to both markers, and restriction analysis of these clones revealed that they can be assembled into maximally four contigs, suggesting that these repetitive probes may be useful for the physical mapping of the Fr locus.
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U2 - 10.1023/A:1006234823105
DO - 10.1023/A:1006234823105
M3 - Article
C2 - 10527422
AN - SCOPUS:0033180242
SN - 0167-4412
VL - 40
SP - 977
EP - 983
JO - Plant molecular biology
JF - Plant molecular biology
IS - 6
ER -