Construction and characterization of a sugar beet (Beta vulgaris) fosmid library

Genome ◽  
2008 ◽  
Vol 51 (11) ◽  
pp. 948-951 ◽  
Author(s):  
Cornelia Lange ◽  
Daniela Holtgräwe ◽  
Britta Schulz ◽  
Bernd Weisshaar ◽  
Heinz Himmelbauer
Keyword(s):  
Sugar Beet ◽  
Beta Vulgaris ◽  
Fosmid Library ◽  
Average Insert Size ◽  
Size Estimation ◽  
Insert Size ◽  
Pcr Screening ◽  
Organellar Dna ◽  
Fosmid End Sequences ◽  
End Sequences

A sugar beet ( Beta vulgaris ) fosmid library from the doubled haploid accession KWS2320 encompassing 115 200 independent clones was constructed and characterized. The average insert size of the fosmid library was determined by pulsed field gel electrophoresis to be 39 kbp on average, thus representing 5.9-fold coverage of the sugar beet genome (758 Mbp). PCR screening of plate pools with primer pairs against nine sugar beet genes supported the insert size estimation. BLAST searches with 2951 fosmid end-sequences originating from 1510 clones (1536 clones attempted) revealed little contamination with organellar DNA (2.1% chloroplast DNA, 0.3% mitochondrial DNA). The sugar beet fosmid library will be integrated in the presently ongoing efforts to determine the sequence of the sugar beet genome. Fosmids will be publicly available in the format of plate pools and individual clones.

Construction and characterization of a BAC library for the molecular dissection of a single wild beet centromere and sugar beet (Beta vulgaris) genome analysis

Genome ◽  
2001 ◽  
Vol 44 (5) ◽  
pp. 846-855 ◽  
Author(s):  
Frank Gindullis ◽  
Daryna Dechyeva ◽  
Thomas Schmidt
Keyword(s):  
Sugar Beet ◽  
Beta Vulgaris ◽  
Bac Library ◽  
Artificial Chromosome ◽  
Average Insert Size ◽  
Insert Size ◽  
Single Chromosome ◽  
Beta Procumbens ◽  
Wild Beet

We have constructed a sugar beet bacterial artificial chromosome (BAC) library of the chromosome mutant PRO1. This Beta vulgaris mutant carries a single chromosome fragment of 6-9 Mbp that is derived from the wild beet Beta procumbens and is transmitted efficiently in meiosis and mitosis. The library consists of 50 304 clones, with an average insert size of 125 kb. Filter hybridizations revealed that approximately 3.1% of the clones contain mitochondrial or chloroplast DNA. Based on a haploid genome size of 758 Mbp, the library represents eight genome equivalents. Thus, there is a greater than 99.96% probability that any sequence of the PRO1 genome can be found in the library. Approximately 0.2% of the clones hybridized with centromeric sequences of the PRO1 minichromosome. Using the identified BAC clones in fluorescence in situ hybridization experiments with PRO1 and B. procumbens chromosome spreads, their wild-beet origin and centromeric localization were demonstrated. Comparative Southern hybridization of pulsed-field separated PRO1 DNA and BAC inserts indicate that the centromeric region of the minichromosome is represented by overlapping clones in the library. Therefore, the PRO1 BAC library provides a useful tool for the characterization of a single plant centromere and is a valuable resource for sugar beet genome analysis.Key words: Beta vulgaris, BAC library, Beta procumbens minichromosome, centromere, FISH.

scholarly journals Construction, Characterization, and Preliminary BAC-End Sequence Analysis of a Bacterial Artificial Chromosome Library of the Tea Plant (Camellia sinensis)

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Jinke Lin ◽  
Dave Kudrna ◽  
Rod A. Wing
Keyword(s):  
Camellia Sinensis ◽  
Bac Library ◽  
Gc Content ◽  
Artificial Chromosome ◽  
Tea Plant ◽  
Average Insert Size ◽  
Plant Genomics ◽  
Insert Size ◽  
Sequence Class ◽  
End Sequences

We describe the construction and characterization of a publicly available BAC library for the tea plant,Camellia sinensis. Using modified methods, the library was constructed with the aim of developing public molecular resources to advance tea plant genomics research. The library consists of a total of 401,280 clones with an average insert size of 135 kb, providing an approximate coverage of 13.5 haploid genome equivalents. No empty vector clones were observed in a random sampling of 576 BAC clones. Further analysis of 182 BAC-end sequences from randomly selected clones revealed a GC content of 40.35% and low chloroplast and mitochondrial contamination. Repetitive sequence analyses indicated that LTR retrotransposons were the most predominant sequence class (86.93%–87.24%), followed by DNA retrotransposons (11.16%–11.69%). Additionally, we found 25 simple sequence repeats (SSRs) that could potentially be used as genetic markers.

Construction of a Coix BAC library and isolation of the 22 kDa α-coixin gene cluster

Genome ◽  
2010 ◽  
Vol 53 (9) ◽  
pp. 667-674 ◽  
Author(s):  
Xiangzong Meng ◽  
Binbin Huang ◽  
Liangliang Zhou ◽  
Yunxia He ◽  
Qi Chen ◽  
...  
Keyword(s):  
Genetic Resource ◽  
Crop Improvement ◽  
Bac Library ◽  
Artificial Chromosome ◽  
Gene Isolation ◽  
Close Relative ◽  
Average Insert Size ◽  
Insert Size ◽  
Bac Clones ◽  
Organellar Dna

Coix lacryma-jobi L. (Coix) is a close relative of maize and is considered a valuable genetic resource for crop improvement. Here we report the construction of the first Coix bacterial artificial chromosome (BAC) library using accession PI 324059. This BAC library contains about 230 400 clones with an average insert size of 113 kb, has low organellar DNA contamination, and provides 16.3-fold coverage of the genome. The library was stored in 12 × 96 pools that could be screened with a PCR protocol. Library screening was performed for the 22 kDa α-coixin gene family. A total of 57 positive pools were identified, and single clones were isolated from 19 of these pools. Based on DNA fingerprinting and Southern blot analysis, these 19 BAC clones form a single contig of about 340 kb in length, indicating that the 22 kDa α-coixin genes occur in a cluster. These results demonstrated the suitability of this BAC library for gene isolation and comparative genomics studies of the Coix genome.

scholarly journals Construction of a BAC Library Covering a Common Region between Two Sugar Beet Translocation Lines with Two Cyst Nematode Resistance Genes

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 822C-822
Author(s):  
Suren Samuelian*
Keyword(s):  
Sugar Beet ◽  
Bac Library ◽  
Artificial Chromosome ◽  
Nematode Resistance ◽  
Cyst Nematode ◽  
Translocation Line ◽  
Average Insert Size ◽  
Insert Size ◽  
Specific Sequence ◽  
Wild Beet

Resistance against the beet cyst nematode (BCN) has been introduced into cultivated sugar beet from wild beet by conventional breeding. The first gene effective against the BCN, Hs1pro-1, was isolated from the sugar beet translocation line A906001. It is assumed that a second nematode resistance gene, Hs1pro-1, is present in the translocation line PRO3, which does not carry Hs1pro-1 but still imparts complete resistance against the nematode and resides in the overlapping region between the two lines. The overall goal of this study was to construct a bacterial artificial chromosome (BAC) library to facilitate the cloning of Hs1pro-1. A BAC library from PRO3 was constructed containing 45,041 clones with an average insert size of 108.36 kb. Screening of the library with organelle specific probes indicated less than 1% mitochondrial and 4% chloroplast DNA content. The library covers 6.17 genome equivalents which provides a 99.76% probability of recovering any specific sequence present in the genome.

Plant regeneration from protoplasts of sugar beet (Beta vulgaris)

1995 ◽  
Vol 94 (2) ◽  
pp. 342-350 ◽  
Author(s):  
Steffen Lenzner ◽  
Kurt Zoglauer ◽  
Otto Schieder
Keyword(s):  
Plant Regeneration ◽  
Sugar Beet ◽  
Beta Vulgaris
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