Frontiers of Agriculture in China >
PCR-based screening of BAC clones of different chromosomes in Chinese cabbage
Received date: 05 Mar 2011
Accepted date: 01 May 2011
Published date: 05 Dec 2011
Copyright
In this paper, taking SSR and functional gene sequence as the primers and the plasmid of first- and second-level pools of bacterial artificial chromosome (BAC) library as templates, the PCR method was used for specific clones of different chromosomes in Chinese cabbage. The results showed that the number of positive clones was 1–11 per primer and the average number of clone was 3.9 by screening 19200 clones of BAC library using 12 pairs of SSR primers from 10 linkage groups individually, which were nearly consistent with about 3.4 times of genome coverage. Positive clones were acquired in chromosome Nos. 2 to 5 and 8 to 10 without screening with the positive clones in chromosome Nos. 1, 6, and 7. In addition, the primer of FLC1 functional gene of chromosome No. 10 was used for PCR screening, and two BAC clones containing FLC1 gene were acquired. Therefore, different specific BAC clones of chromosomes were taken by using SSR primer and functional gene primer. Specific clone screening of chromosomes could provide a probe for identifying the chromosome accurately. Meanwhile, the BAC library screening method was optimized, serving as an effective technical means for quick BAC clone screening.
Key words： Chinese cabbage; chromosome; SSR primer; mixing pool; PCR screening
Daling FENG , Shuxin XUAN , Aixia GU , Airu MA , Jiuhuan LI , Shuxing SHEN . PCR-based screening of BAC clones of different chromosomes in Chinese cabbage[J]. Frontiers of Agriculture in China, 2011 , 5(4) : 524 -528 . DOI: 10.1007/s11703-011-1122-2
| 1 |
Achenbach U C, Tang X, Ballvora A, de Jong H, Gebhardt C (2010). Comparison of the chromosome maps around a resistance hot spot on chromosome 5 of potato and tomato using BAC-FISH painting. Genome, 53(2): 103-110
|
| 2 |
Febrer M, Cheung F, Town C D, Cannon S B, Young N D, Abberton M T, Jenkins G, Milbourne D (2007). Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.). Genome, 50(4): 412-421
|
| 3 |
Giraudat J, Hauge B M, Valon C, Smalle J, Parcy F, Goodman H M (1992). Isolation of the Arabidopsis ABI3 gene by positional cloning. Plant Cell, 4(10): 1251-1261
|
| 4 |
Guan Y, Chen Q, Pan J S, Li Z, He H L, Wu A Z, Song R T, Cai R (2008). Isolation of linkage special clones and construction of BAC library in cucumber (Cucumis sativus L.). Progress in Natural Science, 18(2): 211-215 (in Chinese)
|
| 5 |
Guan Y, Chen Q, Pan J S, Li Z, He H L, Wu A Z, Song R T, Cai R (2008). BAC library building and screening of linkage specific clones on the study of Cucumis sativus L.. Prog Nat Sci, 18(2): 211-215 (in Chinese)
|
| 6 |
Huan P, Zhang X J, Li F H, Zhang Y, Zhao C, Liu B Z, Xiang J H (2009). Chromosomal localization and development of SNP markers of a serine protease gene in Farrer’s scallop (Chlamys farreri). Hereditas, 31(12): 1241-1247 (in Chinese)
|
| 7 |
Li X J (2010). Establishment and SSR identification of a few cabbage-Chinese cabbage alien addition lines. Dissertation for the Master Degree. Baoding: Agricultural University of Hebei, 24
|
| 8 |
Liu H B, Liu K, Wang J F, Ma R Z (2006). A BAC clone-based physical map of ovine major histocompatibility complex. Genomics, 88(1): 88-95
|
| 9 |
Luo R, Wu W L, Zhang Y, Li Y H (2010). SSR Marker and its application to crop genetics and breeding. Genomics and Applied Biology, 29(1): 137-143 (in Chinese)
|
| 10 |
Mun J H, Kwon S J, Yang T J, Kim H S, Choi B S, Baek S, Kim J S, Jin M, Kim J A, Lim M H, Lee S I, Kim H I, Kim H, Lim Y P, Park B S (2008). The first generation of a BAC-based physical map of Brassica rapa. BMC Genomics, 9: 280
|
| 11 |
Osoegawa K, Woon P Y, Zhao B, Frengen E, Tateno M, Catanese J J, de Jong P J (1998). An improved approach for construction of bacterial artificial chromosome libraries. Genomics, 52(1): 1-8
|
| 12 |
Schranz M E, Quijada P, Sung S B, Lukens L, Amasino R, Osborn T C (2002). Characterization and effects of the replicated flowering time gene FLC in Brassica rapa. Genetics, 162(3): 1457-1468
|
| 13 |
Shizuya H, Birren B, Kim U J, Mancino V, Slepak T, Tachiiri Y, Simon M (1992). Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proc Natl Acad Sci USA, 89(18): 8794-8797
|
| 14 |
Szinay D, Chang S B, Khrustaleva L, Peters S, Schijlen E, Bai Y, Stiekema W J, van Ham R C, de Jong H, Klein Lankhorst R M (2008). High-resolution chromosome mapping of BACs using multi-colour FISH and pooled-BAC FISH as a backbone for sequencing tomato chromosome 6. Plant J, 56(4): 627-637
|
| 15 |
Terol J, Naranjo M A, Ollitrault P, Talon M (2008). Development of genomic resources for Citrus clementina: Characterization of three deep-coverage BAC libraries and analysis of 46,000 BAC end sequences. BioMed Central Genomics, 9: 423
|
| 16 |
Xiong Z Y, Tan G X, You A Q, He G Y, She C W, Li L J, Song Y C (2004). Comparative physical positioning of BAC cloning linked with Glh, Bph-3 and xa-5 in cultivated rice and Oryza granulate. Chin Sci Bull, 49(3): 252-257 (in Chinese)
|
/
| 〈 |
|
〉 |