Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137
BioMed research international, Vol.2014(2014), pp.1-9
2014
:
https://hdl.handle.net/2376/118122
: PMC4026951
: 24895618
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.
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- Construction and characterization of a bacterial artificial chromosome library for the hexaploid wheat line 92R137
- Qingdong Zeng - Northwest A&F UniversityXin Xu - Northwest A&F UniversityXue Shi - State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, ChinaXiaojun Nie - State Key Laboratory of Crop Stress Biology for Arid Areas and College of Agronomy, Northwest A&F University, Yangling, Shaanxi 712100, ChinaHua Zhuang - State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, ChinaXianming Chen - 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, USAXiaojie Wang - Northwest A&F UniversityLili Huang - Northwest A&F UniversityZhensheng Kang - State Key Laboratory of Crop Stress Biology for Arid Areas and College of Plant Protection, Northwest A&F University, Yangling, Shaanxi 712100, ChinaFengping Yuan - Northwest A&F UniversityZhonghua Wang - Northwest A&F UniversityDejun Han - Northwest A&F University
- BioMed research international, Vol.2014(2014), pp.1-9
- Department of Plant Pathology
- Hindawi Publishing Corporation; United States
- 31371924 / http://dx.doi.org/10.13039/501100001809 National Natural Science Foundation of China 2011ZX08002-001 / National Major Project of Breeding for New Transgenic Organisms in China B07049 / http://dx.doi.org/10.13039/501100002338 Ministry of Education of the People's Republic of China 2013CB127700 / National Basic Research Program of China
- 99900547803801842
- English
- Journal article