Construction of BAC and BIBAC libraries from sunflower and identification of linkage group-specific clones by overgo hybridization.

TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik
Jiuhuan FengC C Jan

Abstract

Complementary BAC and BIBAC libraries were constructed from nuclear DNA of sunflower cultivar HA 89. The BAC library, constructed with BamHI in the pECBAC1 vector, contains 107,136 clones and has an average insert size of 140 kb. The BIBAC library was constructed with HindIII in the plant-transformation-competent binary vector pCLD04541 and contains 84,864 clones, with an average insert size of 137 kb. The two libraries combined contain 192,000 clones and are equivalent to approximately 8.9 haploid genomes of sunflower (3,000 Mb/1C), and provide a greater than 99% probability of obtaining a clone of interest. The frequencies of BAC and BIBAC clones carrying chloroplast or mitochondrial DNA sequences were estimated to be 2.35 and 0.04%, respectively, and insert-empty clones were less than 0.5%. To facilitate chromosome engineering and anchor the sunflower genetic map to its chromosomes, one to three single- or low-copy RFLP markers from each linkage group of sunflower were used to design pairs of overlapping oligonucleotides (overgos). Thirty-six overgos were designed and pooled as probes to screen a subset (5.1x) of the BAC and BIBAC libraries. Of the 36 overgos, 33 (92%) gave at least one positive clone and 3 (8%) failed to hi...Continue Reading

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Citations

Apr 15, 2006·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Junfang ChenC C Jan
Apr 26, 2008·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Jiuhuan Feng, Chao-Chien Jan
Jun 27, 2009·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Sujatha MulpuriChao-Chien Jan
Feb 22, 2012·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Zhao LiuChao-Chien Jan
Jul 2, 2011·Plant Methods·Wilma SabettaCinzia Montemurro
Jan 15, 2013·G3 : Genes - Genomes - Genetics·Jiuhuan FengChao-Chien Jan
Jul 28, 2018·Plant Molecular Biology·Jing ZhouMeng-Zhu Lu

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