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. 2011:2011:476723.
doi: 10.1155/2011/476723. Epub 2010 Dec 23.

Construction, characterization, and preliminary BAC-end sequence analysis of a bacterial artificial chromosome library of the tea plant (Camellia sinensis)

Jinke Lin  1 , Dave Kudrna, Rod A Wing
Affiliations

Construction, characterization, and preliminary BAC-end sequence analysis of a bacterial artificial chromosome library of the tea plant (Camellia sinensis)

Jinke Lin et al. J Biomed Biotechnol. 2011.

Abstract

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.

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Figures

Figure 1
Figure 1
Insert DNA analysis of random BAC clones from the Camellia sinensis HindIII BAC library, CSBCBa, by pulsed-field gel electrophoresis. DNA samples were digested with NotI and separated by pulsed-field gel electrophoresis with ramp pulse time of 5–15 s at 6 V/cm at 14°C in 0.5× TBE buffer for 16 h. Markers used are midrange I (outside lanes). The 7.5-kb common band corresponds to the cloning vector pIndigoBAC536 SwaI (pAGIBAC1).
Figure 2
Figure 2
Insert size distribution of Camellia sinensis BAC clones from the CSBCBa library. The average insert size of the BAC library was calculated to be 135 kb from an analyzed random sample size of 576 BAC clones.
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