Construction and Characterization of a Bacterial Artificial Chromosome Library for the Hexaploid Wheat Line 92R137

For map-based cloning of genes conferring important traits in the hexaploid wheat line 92R137, a bacterial artificial chromosome (BAC) library, including two sublibraries, was constructed using the genomic DNA of 92R137 digested with restriction enzymesHindIII andBamHI. The BAC library was composed of total 765,696 clones, of which 390,144 were from theHindIII digestion and 375,552 from theBamHI digestion. Through pulsed-field gel electrophoresis (PFGE) analysis of 453 clones randomly selected from theHindIII sublibrary and 573 clones from theBamHI sublibrary, the average insert sizes were estimated as 129 and 113 kb, respectively. Thus, theHindIII sublibrary was estimated to have a 3.01-fold coverage and theBamHI sublibrary a 2.53-fold coverage based on the estimated hexaploid wheat genome size of 16,700 Mb. The 765,696 clones were arrayed in 1,994 384-well plates. All clones were also arranged into plate pools and further arranged into 5-dimensional (5D) pools. The probability of identifying a clone corresponding to any wheat DNA sequence (such as geneYr26for stripe rust resistance) from the library was estimated to be more than 99.6%. Through polymerase chain reaction screening the 5D pools withXwe173, a marker tightly linked toYr26, six BAC clones were successfully obtained. These results demonstrate that the BAC library is a valuable genomic resource for positional cloning ofYr26and other genes of interest.

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Construction of a hexaploid wheat (Triticum aestivumL.) bacterial artificial chromosome library for cloning genes for stripe rust resistance

Genome ◽

10.1139/g05-078 ◽

2005 ◽

Vol 48 (6) ◽

pp. 1028-1036 ◽

Cited By ~ 13

Author(s):

P Ling ◽

X M Chen

Keyword(s):

Triticum Aestivum ◽

Stripe Rust ◽

Hexaploid Wheat ◽

Rust Resistance ◽

Puccinia Striiformis ◽

Bac Library ◽

Artificial Chromosome ◽

Stripe Rust Resistance ◽

Insert Size ◽

Bac Clones

A hexaploid wheat (Triticum aestivum L.) bacterial artificial chromosome (BAC) library was constructed for cloning Yr5 and other genes conferring resistance to stripe rust (Puccinia striiformis f. sp. tritici). Intact nuclei from a Yr5 near-isogenic line were used to isolate high molecular weight DNA, which was partially cleaved with HindIII and cloned into pECBAC1 and pIndigoBAC-5 vectors. The wheat BAC library consisted of 422 400 clones arrayed in 1100 micro-titer plates (each plate with 384 wells). Random sampling of 300 BAC clones indicated an average insert size of 140 kb, with a size range from 25 to 365 kb. Ninety percent of the clones in the library had an insert size greater than 100 kb and fewer than 5% of the clones did not contain inserts. Based on an estimated genome size of 15 966 Mb for hexaploid wheat, the BAC library was estimated to have a total coverage of 3.58× wheat genome equivalents, giving approximately 96% probability of identifying a clone representing any given wheat DNA sequence. Twelve BAC clones containing an Yr5 locus-specific marker (Yr5STS7/8) were successfully selected by PCR screening of 3-dimensional BAC pools. The results demonstrated that the T. aestivum BAC library is a valuable genomic resource for positional cloning of Yr5. The library also should be useful in cloning other genes for stripe rust resistance and other traits of interest in hexaploid wheat.Key words: BAC library, BAC pools, hexaploid wheat, Puccinia striiformis f. sp. tritici, resistance gene, stripe rust, Triticum aestivum.

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Construction and Characterization of a Bacterial Artificial Chromosome (BAC) Library from the Japanese Malting Barley variety ‘Haruna Nijo’

Breeding Science ◽

10.1270/jsbbs.57.29 ◽

2007 ◽

Vol 57 (1) ◽

pp. 29-38 ◽

Cited By ~ 35

Author(s):

Daisuke Saisho ◽

Eriko Myoraku ◽

Shinji Kawasaki ◽

Kazuhiro Sato ◽

Kazuyoshi Takeda

Keyword(s):

Bacterial Artificial Chromosome ◽

Bac Library ◽

Artificial Chromosome ◽

Barley Variety ◽

Malting Barley

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Construction and characterization of the first bacterial artificial chromosome library for the cotton species Gossypium barbadense L.

Genome ◽

10.1139/g06-113 ◽

2006 ◽

Vol 49 (11) ◽

pp. 1393-1398 ◽

Cited By ~ 12

Author(s):

X.F. Wang ◽

J. Ma ◽

W.S. Wang ◽

Y.M. Zheng ◽

G.Y. Zhang ◽

...

Keyword(s):

Bacterial Artificial Chromosome ◽

Bac Library ◽

Artificial Chromosome ◽

Gossypium Barbadense ◽

Average Insert Size ◽

Insert Size ◽

Breeding Programs ◽

Fiber Properties ◽

Cotton Species

As the second most widely cultivated cotton, Gossypium barbadense is well known for its superior fiber properties and its high levels of resistance to Fusarium and Verticillium wilts. To enhance our ability to exploit these properties in breeding programs, we constructed the first bacterial artificial chromosome (BAC) library for this species. The library contains 167 424 clones (49 920 BamHI and 117 504 HindIII clones), with an estimated average insert size of 130 kb. About 94.0% of the clones had inserts over 100 kb, and the empty clones accounted for less than 4.0%. Contamination of the library with chloroplast clones was very low (0.2%). Screening the library with locus-specific probes showed that BAC clones represent 6.5-fold genome equivalents. This high-quality library provides an additional asset with which to exploit genetic variation for cotton improvement.

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Bacterial artificial chromosome (BAC) library resource for positional cloning of pest and disease resistance genes in cassava (Manihot esculenta Crantz)

Plant Molecular Biology ◽

10.1007/s11103-004-5045-7 ◽

2004 ◽

Vol 56 (4) ◽

pp. 555-561 ◽

Cited By ~ 16

Author(s):

J. Tomkins ◽

M. Fregene ◽

D. Main ◽

H. Kim ◽

R. Wing ◽

...

Keyword(s):

Disease Resistance ◽

Bacterial Artificial Chromosome ◽

Resistance Genes ◽

Manihot Esculenta ◽

Positional Cloning ◽

Bac Library ◽

Artificial Chromosome ◽

Disease Resistance Genes ◽

Manihot Esculenta Crantz ◽

Library Resource

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Construction and Characterization of a Bacterial Artificial Chromosome (BAC) Library of Pacific White Shrimp, Litopenaeus vannamei

Marine Biotechnology ◽

10.1007/s10126-009-9209-y ◽

2009 ◽

Vol 12 (2) ◽

pp. 141-149 ◽

Cited By ~ 21

Author(s):

Xiaojun Zhang ◽

Yang Zhang ◽

Chantel Scheuring ◽

Hong-Bin Zhang ◽

Pin Huan ◽

...

Keyword(s):

Bacterial Artificial Chromosome ◽

Litopenaeus Vannamei ◽

Bac Library ◽

Artificial Chromosome ◽

Pacific White Shrimp ◽

White Shrimp

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Construction and Characterization of a Bacterial Artificial Chromosome Library for the A-Genome of Cotton (G.arboreumL.)

Journal of Biomedicine and Biotechnology ◽

10.1155/2011/457137 ◽

2011 ◽

Vol 2011 ◽

pp. 1-5 ◽

Cited By ~ 5

Author(s):

Yan Hu ◽

Yamin Lu ◽

Dan Ma ◽

Wangzhen Guo ◽

Tianzhen Zhang

Keyword(s):

Bacterial Artificial Chromosome ◽

Bac Library ◽

Artificial Chromosome ◽

Gene Isolation ◽

Comparative Genome Analysis ◽

Bac Clones ◽

A Genome ◽

Reference Map ◽

Insert Length

A bacterial artificial chromosome (BAC) library for the A-genome of cotton has been constructed from the leaves ofG. arboreumL cv. Jianglinzhongmian. It is used as elite A-genome germplasm resources in the present cotton breeding program and has been used to build a genetic reference map of cotton. The BAC library consists of 123,648 clones stored in 322 384-well plates. Statistical analysis of a set of 103 randomly selected BAC clones indicated that each clone has an average insert length of 100.2 kb per plasmid, with a range of 30 to 190 kb. Theoretically, this represents 7.2 haploid genome equivalents based on an A-genome size of 1697 Mb. The BAC library has been arranged in column pools and superpools allowing screening with various PCR-based markers. In the future, the A-genome cotton BAC library will serve as both a giant gene resource and a valuable tool for map-based gene isolation, physical mapping and comparative genome analysis.

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Characterization of an Aedes aegypti bacterial artificial chromosome (BAC) library and chromosomal assignment of BAC clones for physical mapping quantitative trait loci that influence Plasmodium susceptibility

Insect Molecular Biology ◽

10.1046/j.0962-1075.2004.00456.x ◽

2004 ◽

Vol 13 (1) ◽

pp. 37-44 ◽

Cited By ~ 19

Author(s):

L. V. Jiménez ◽

B.-K. Kang ◽

B. DeBruyn ◽

D. D. Lovin ◽

D. W. Severson

Keyword(s):

Quantitative Trait Loci ◽

Bacterial Artificial Chromosome ◽

Aedes Aegypti ◽

Physical Mapping ◽

Quantitative Trait ◽

Bac Library ◽

Artificial Chromosome ◽

Chromosomal Assignment ◽

Bac Clones

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Construction of a marsupial bacterial artificial chromosome library from the model Australian marsupial, the tammar wallaby (Macropus eugenii)

Australian Journal of Zoology ◽

10.1071/zo05033 ◽

2005 ◽

Vol 53 (6) ◽

pp. 389 ◽

Cited By ~ 6

Author(s):

Natasha Sankovic ◽

Wayne Bawden ◽

John Martyn ◽

Jennifer A. M. Graves ◽

Kurt Zuelke

Keyword(s):

Bacterial Artificial Chromosome ◽

Positional Cloning ◽

Bac Library ◽

Artificial Chromosome ◽

Single Copy ◽

Tammar Wallaby ◽

Average Insert Size ◽

Insert Size ◽

Macropus Eugenii ◽

Australian Marsupial

With the accelerating recognition of the power of comparative genomics, there is now enormous interest in sequencing the genomes of a broad range of species. Marsupials diverged at an important evolutionary time. The model Australian marsupial, the tammar wallaby (Macropus eugenii), has long been a resource for biological and genetic studies of marsupials, and the availability of a bacterial artificial chromosome (BAC) library will be a valuable resource in these studies. A tammar wallaby BAC library was constructed using pRazorBAC vector. It contains 55 296 clones with an average insert size of 108 kb, representing 2.2 times coverage of the wallaby genome (based on an estimated 2.7 × 109 bp haploid genome size). The library was arrayed in 384-well plates, and spotted in duplicate onto nylon membranes. Screening these membranes has yielded clones containing 34 single-copy genes distributed over the genome, while it failed for only one gene. Each probe isolated 1–12 BAC clones and, to date, no chimeric clones have been found. This BAC library will constitute an invaluable resource for creating physical maps, positional cloning of genes and other sequences in the tammar wallaby, as well as comparative mapping studies in mammals.

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