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Microdissection of the Ah01 chromosome in upland cotton and microcloning of resistance gene anologs from the single chromosome.
Wednesday, 2017/06/07 | 08:12:14

Cao X, Liu Y, Liu Z, Liu F, Wu Y, Zhou Z, Cai X, Wang X, Zhang Z, Wang Y, Luo Z, Peng R, Wang K.

Hereditas. 2017 May 18;154:13. doi: 10.1186/s41065-017-0035-3. eCollection 2017.

Abstract

BACKGROUND:

Chromosome microdissection is one of the most important techniques in molecular cytogenetic research. Cotton (Gossypium Linnaeus, 1753) is the main natural fiber crop in the world. The resistance gene analog (RGA) cloning after its single chromosome microdissection can greatly promote cotton genome research and breeding.

RESULTS:

Using the linker adaptor PCR (LA-PCR) with the primers of rice disease-resistance homologues, three nucleotide sequences PS016 (KU051681), PS054 (KU051682), and PS157 (KU051680) were obtained from the chromosome Ah01 of upland cotton (cv. TM-1). The Blast results showed that the three sequences are the nucleotide binding site-leucine rich repeat (NBS-LRR) type RGAs. Clustering results indicated that they are homologous to these published RGAs. Thus, the three RGAs can definitely be confirmed as NBS-LRR class of RGAs in upland cotton.

CONCLUSIONS:

Using single chromosome microdissection technique, DNA libraries containing cotton RGAs were obtained. This technique can promote cotton gene cloning, marker development and even the improvement of cotton genome research and breeding.

See https://www.ncbi.nlm.nih.gov/pubmed/28529470

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Object name is 41065_2017_35_Fig2_HTML.jpg

Figure 4: PAGE of SSR primer amplification product from single chromosome pool. 19: SSR primer amplification products from partial other chromosomes pool with Ah01 chromosome specific primer. 10: SSR primer amplification products from single chromosome pool with chromosome Ah01 special primer (arrow indicated). 11: The negative control. 12: The positive control. M: DNA marker

 

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