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Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137.
Biomed Res Int. 2014; 2014:845806.BR

Abstract

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 enzymes HindIII and BamHI. The BAC library was composed of total 765,696 clones, of which 390,144 were from the HindIII digestion and 375,552 from the BamHI digestion. Through pulsed-field gel electrophoresis (PFGE) analysis of 453 clones randomly selected from the HindIII sublibrary and 573 clones from the BamHI sublibrary, the average insert sizes were estimated as 129 and 113 kb, respectively. Thus, the HindIII sublibrary was estimated to have a 3.01-fold coverage and the BamHI 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 gene Yr26 for stripe rust resistance) from the library was estimated to be more than 99.6%. Through polymerase chain reaction screening the 5D pools with Xwe173, a marker tightly linked to Yr26, six BAC clones were successfully obtained. These results demonstrate that the BAC library is a valuable genomic resource for positional cloning of Yr26 and other genes of interest.

Authors+Show Affiliations

State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.Wheat Genetics, Quality, Physiology, and Disease Research Unit, Agricultural Research Service, United States Department of Agriculture, and Department of Plant Pathology, Washington State University, Pullman, WA 99164-6430, USA.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, China.State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, China.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

24895618

Citation

Zeng, Qingdong, et al. "Construction and Characterization of a Bacterial Artificial Chromosome Library for the Hexaploid Wheat Line 92R137." BioMed Research International, vol. 2014, 2014, p. 845806.
Zeng Q, Yuan F, Xu X, et al. Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137. Biomed Res Int. 2014;2014:845806.
Zeng, Q., Yuan, F., Xu, X., Shi, X., Nie, X., Zhuang, H., Chen, X., Wang, Z., Wang, X., Huang, L., Han, D., & Kang, Z. (2014). Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137. BioMed Research International, 2014, 845806. https://doi.org/10.1155/2014/845806
Zeng Q, et al. Construction and Characterization of a Bacterial Artificial Chromosome Library for the Hexaploid Wheat Line 92R137. Biomed Res Int. 2014;2014:845806. PubMed PMID: 24895618.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137. AU - Zeng,Qingdong, AU - Yuan,Fengping, AU - Xu,Xin, AU - Shi,Xue, AU - Nie,Xiaojun, AU - Zhuang,Hua, AU - Chen,Xianming, AU - Wang,Zhonghua, AU - Wang,Xiaojie, AU - Huang,Lili, AU - Han,Dejun, AU - Kang,Zhensheng, Y1 - 2014/05/05/ PY - 2014/01/28/received PY - 2014/03/26/revised PY - 2014/04/18/accepted PY - 2014/6/5/entrez PY - 2014/6/5/pubmed PY - 2015/11/10/medline SP - 845806 EP - 845806 JF - BioMed research international JO - Biomed Res Int VL - 2014 N2 - 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 enzymes HindIII and BamHI. The BAC library was composed of total 765,696 clones, of which 390,144 were from the HindIII digestion and 375,552 from the BamHI digestion. Through pulsed-field gel electrophoresis (PFGE) analysis of 453 clones randomly selected from the HindIII sublibrary and 573 clones from the BamHI sublibrary, the average insert sizes were estimated as 129 and 113 kb, respectively. Thus, the HindIII sublibrary was estimated to have a 3.01-fold coverage and the BamHI 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 gene Yr26 for stripe rust resistance) from the library was estimated to be more than 99.6%. Through polymerase chain reaction screening the 5D pools with Xwe173, a marker tightly linked to Yr26, six BAC clones were successfully obtained. These results demonstrate that the BAC library is a valuable genomic resource for positional cloning of Yr26 and other genes of interest. SN - 2314-6141 UR - https://www.unboundmedicine.com/medline/citation/24895618/Construction_and_characterization_of_a_bacterial_artificial_chromosome_library_for_the_hexaploid_wheat_line_92R137_ DB - PRIME DP - Unbound Medicine ER -