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Analysis of restriction sites and Southern blotting of two molecular markers linked to grape seedless gene

Published online by Cambridge University Press:  12 February 2007

Yang Ke-Qiang
Affiliation:
Key Laboratory of Ministry of Agriculture of Northwest Horticulture Plant Germplasm and Genetic Improvement, College of Horticulture, Northwest A&F University, Yangling 712100, China College of Forestry, Shandong Agricultural University, Taian 271018, China
Wang Yue-Jin*
Affiliation:
Key Laboratory of Ministry of Agriculture of Northwest Horticulture Plant Germplasm and Genetic Improvement, College of Horticulture, Northwest A&F University, Yangling 712100, China
Zhang Jin-Jin
Affiliation:
Key Laboratory of Ministry of Agriculture of Northwest Horticulture Plant Germplasm and Genetic Improvement, College of Horticulture, Northwest A&F University, Yangling 712100, China College of Life Sciences, Shaanxi Normal University, Xi'an 710062, China
Wang Xi-Ping
Affiliation:
Key Laboratory of Ministry of Agriculture of Northwest Horticulture Plant Germplasm and Genetic Improvement, College of Horticulture, Northwest A&F University, Yangling 712100, China
Wan Yi-Zhen
Affiliation:
Key Laboratory of Ministry of Agriculture of Northwest Horticulture Plant Germplasm and Genetic Improvement, College of Horticulture, Northwest A&F University, Yangling 712100, China
Zhang Jian-Xia
Affiliation:
Key Laboratory of Ministry of Agriculture of Northwest Horticulture Plant Germplasm and Genetic Improvement, College of Horticulture, Northwest A&F University, Yangling 712100, China
*
*Corresponding author: Email: wangyj@public.xa.sn.cn

Abstract

The cloning and sequencing of two specific molecular markers linked to grape (Vitis vinifera) seedless gene indicated that the full lengths of the markers, 39970524-5-564 (GenBank accession number: AY327513) and 39970524-6-1538 (GenBank accession number: AY327514), were 564 bp and 1538 bp, respectively. The restriction sites of sequences 39970524-5-564 and 39970524-6-1538 were analysed with Wingene231 software. The results indicated that there were 29 and 130 restriction sites in 39970524-5-564 and 39970524-6-1538, respectively, digested by restriction enzymes with recognition sites of six or more than six bases. There was no cleavage site in 39970524-5-564 and there was one site at 135 bp of 39970524-6-1538 when these two markers were digested by EcoRI. Both markers can not be digested by HindIII. Therefore, the full length of 39970524-5-564 and of the two bands of 39970524-6-1538 (1400 bp and 135 bp) could be cut out from the pGEMR-T Easy Vector by EcoRI. The labelled DNA of 39970524-5-564, used as a probe for Southern blotting with genomic DNA from Red Globe, Flame Seedless, Thompson Seedless, four seedless hybrids and three seeded hybrids, showed a unique hybrid band present in all seedless individuals, but absent in all seeded individuals. It was further indicated that the specific marker 39970524-5-564 was derived from the genome of seedless grapes.

Type
Research Article
Copyright
Copyright © China Agricultural University and Cambridge University Press 2006

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