Generation of a soybean BAC library, and identification of DNA sequences tightly linked to the Rps1-k disease resistance gene

1999 ◽  
Vol 98 (5) ◽  
pp. 712-720 ◽  
Author(s):  
S. S. Salimath ◽  
M. K. Bhattacharyya
1996 ◽  
Vol 14 (2) ◽  
pp. 107-114 ◽  
Author(s):  
Guo-Liang Wang ◽  
Randy Warren ◽  
Goger Innes ◽  
Brian Osborne ◽  
Barbara Baker ◽  
...  

Genome ◽  
1997 ◽  
Vol 40 (4) ◽  
pp. 420-427 ◽  
Author(s):  
L. Fredrick Marek ◽  
R. C. Shoemaker

We constructed a soybean bacterial artificial chromosome (BAC) library suitable for map-based cloning and physical mapping in soybean. This library consists of approximately 40 000 clones (4–5 genome equivalents) stored individually in 384-well microtiter dishes. A random sampling of 224 clones yielded an average insert size of 150 kb, giving a 98% probability of recovering any specific sequence. We screened the library for seven single or very low copy genie or genomic sequences using the polymerase chain reaction (PCR) and found between one and seven BACs for each of the seven sequences. When testing the library with a portion of the soybean psbA chloroplast gene, we found less than 1% chloroplast DNA representation. We also screened the library for eight different classes of disease resistance gene analogs (RGAs) and identified BACs containing all RGAs except class 8. We arranged nine of the class 1 RGA BACs and six of the class 3 RGA BACs into individual contigs based on fingerprint patterns observed after Southern probing of restriction digests of the member BACs with a class-specific sequence. This resulted in the partial localization of the different multigene family sequences without precise definition of their exact positions. Using PCR-based end rescue techniques and RFLP mapping of BAC ends, we mapped individual BACs of each contig onto linkage group J of the soybean public map. The class 1 contig mapped to the region on linkage group J that contains several disease resistance genes. The class 1 contig extended approximately 400 kb. The arrangement of the BACs within this contig has been confirmed using PCR. One end of the class 1 contig core BAC mapped to two positions on linkage group J and cosegregated with two class 1 RGA loci, suggesting that this segment is within an area of regional duplication.Key words: bacterial artificial chromosome, BAC library, soybean, contig, resistance gene analog.


2012 ◽  
Vol 34 (1) ◽  
pp. 56
Author(s):  
Ling CHEN ◽  
Hao ZHANG ◽  
Xian-Qin QIU ◽  
Hui-Jun YAN ◽  
Qi-Gang WANG ◽  
...  

Genetics ◽  
2002 ◽  
Vol 162 (4) ◽  
pp. 1961-1977
Author(s):  
Michelle A Graham ◽  
Laura Fredrick Marek ◽  
Randy C Shoemaker

Abstract PCR amplification was previously used to identify a cluster of resistance gene analogues (RGAs) on soybean linkage group J. Resistance to powdery mildew (Rmd-c), Phytophthora stem and root rot (Rps2), and an ineffective nodulation gene (Rj2) map within this cluster. BAC fingerprinting and RGA-specific primers were used to develop a contig of BAC clones spanning this region in cultivar “Williams 82” [rps2, Rmd (adult onset), rj2]. Two cDNAs with homology to the TIR/NBD/LRR family of R-genes have also been mapped to opposite ends of a BAC in the contig Gm_Isb001_091F11 (BAC 91F11). Sequence analyses of BAC 91F11 identified 16 different resistance-like gene (RLG) sequences with homology to the TIR/NBD/LRR family of disease resistance genes. Four of these RLGs represent two potentially novel classes of disease resistance genes: TIR/NBD domains fused inframe to a putative defense-related protein (NtPRp27-like) and TIR domains fused inframe to soybean calmodulin Ca2+-binding domains. RT-PCR analyses using gene-specific primers allowed us to monitor the expression of individual genes in different tissues and developmental stages. Three genes appeared to be constitutively expressed, while three were differentially expressed. Analyses of the R-genes within this BAC suggest that R-gene evolution in soybean is a complex and dynamic process.


BMC Genomics ◽  
2011 ◽  
Vol 12 (1) ◽  
Author(s):  
Alessandra F Ribas ◽  
Alberto Cenci ◽  
Marie-Christine Combes ◽  
Hervé Etienne ◽  
Philippe Lashermes

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