Characterization of an Aedes aegypti bacterial artificial chromosome (BAC) library and chromosomal assignment of BAC clones for physical mapping quantitative trait loci that influence Plasmodium susceptibility

2004 ◽  
Vol 13 (1) ◽  
pp. 37-44 ◽  
Author(s):  
L. V. Jiménez ◽  
B.-K. Kang ◽  
B. DeBruyn ◽  
D. D. Lovin ◽  
D. W. Severson
2011 ◽  
Vol 2011 ◽  
pp. 1-5 ◽  
Author(s):  
Yan Hu ◽  
Yamin Lu ◽  
Dan Ma ◽  
Wangzhen Guo ◽  
Tianzhen Zhang

A bacterial artificial chromosome (BAC) library for the A-genome of cotton has been constructed from the leaves ofG. arboreumL cv. Jianglinzhongmian. It is used as elite A-genome germplasm resources in the present cotton breeding program and has been used to build a genetic reference map of cotton. The BAC library consists of 123,648 clones stored in 322 384-well plates. Statistical analysis of a set of 103 randomly selected BAC clones indicated that each clone has an average insert length of 100.2 kb per plasmid, with a range of 30 to 190 kb. Theoretically, this represents 7.2 haploid genome equivalents based on an A-genome size of 1697 Mb. The BAC library has been arranged in column pools and superpools allowing screening with various PCR-based markers. In the future, the A-genome cotton BAC library will serve as both a giant gene resource and a valuable tool for map-based gene isolation, physical mapping and comparative genome analysis.


Genome ◽  
2004 ◽  
Vol 47 (2) ◽  
pp. 239-245 ◽  
Author(s):  
Yaping Qian ◽  
Li Jin ◽  
Bing Su

The large-insert genomic DNA library is a critical resource for genome-wide genetic dissection of target species. We constructed a high-redundancy bacterial artificial chromosome (BAC) library of a New World monkey species, the black-handed spider monkey (Ateles geoffroyi). A total of 193 152 BAC clones were generated in this library. The average insert size of the BAC clones was estimated to be 184.6 kb with the small inserts (50-100 kb) accounting for less than 3% and the non-recombinant clones only 1.2%. Assuming a similar genome size with humans, the spider monkey BAC library has about 11× genome coverage. In addition, by end sequencing of randomly selected BAC clones, we generated 367 sequence tags for the library. When blasted against human genome, they showed a good correlation between the number of hit clones and the size of the chromosomes, an indication of unbiased chromosomal distribution of the library. This black-handed spider monkey BAC library would serve as a valuable resource in comparative genomic study and large-scale genome sequencing of nonhuman primates.Key words: black-handed spider monkeys, Ateles geoffroyi, BAC library.


1998 ◽  
Vol 66 (5) ◽  
pp. 2221-2229 ◽  
Author(s):  
Roland Brosch ◽  
Stephen V. Gordon ◽  
Alain Billault ◽  
Thierry Garnier ◽  
Karin Eiglmeier ◽  
...  

ABSTRACT The bacterial artificial chromosome (BAC) cloning system is capable of stably propagating large, complex DNA inserts in Escherichia coli. As part of the Mycobacterium tuberculosis H37Rv genome sequencing project, a BAC library was constructed in the pBeloBAC11 vector and used for genome mapping, confirmation of sequence assembly, and sequencing. The library contains about 5,000 BAC clones, with inserts ranging in size from 25 to 104 kb, representing theoretically a 70-fold coverage of the M. tuberculosisgenome (4.4 Mb). A total of 840 sequences from the T7 and SP6 termini of 420 BACs were determined and compared to those of a partial genomic database. These sequences showed excellent correlation between the estimated sizes and positions of the BAC clones and the sizes and positions of previously sequenced cosmids and the resulting contigs. Many BAC clones represent linking clones between sequenced cosmids, allowing full coverage of the H37Rv chromosome, and they are now being shotgun sequenced in the framework of the H37Rv sequencing project. Also, no chimeric, deleted, or rearranged BAC clones were detected, which was of major importance for the correct mapping and assembly of the H37Rv sequence. The minimal overlapping set contains 68 unique BAC clones and spans the whole H37Rv chromosome with the exception of a single gap of ∼150 kb. As a postgenomic application, the canonical BAC set was used in a comparative study to reveal chromosomal polymorphisms between M. tuberculosis, M. bovis, and M. bovis BCG Pasteur, and a novel 12.7-kb segment present in M. tuberculosis but absent from M. bovis and M. bovis BCG was characterized. This region contains a set of genes whose products show low similarity to proteins involved in polysaccharide biosynthesis. The H37Rv BAC library therefore provides us with a powerful tool both for the generation and confirmation of sequence data as well as for comparative genomics and other postgenomic applications. It represents a major resource for present and future M. tuberculosis research projects.


2011 ◽  
Vol 92 (7) ◽  
pp. 1500-1507 ◽  
Author(s):  
Stephen J. Spatz ◽  
Lorraine P. Smith ◽  
Susan J. Baigent ◽  
Lawrence Petherbridge ◽  
Venugopal Nair

The identification of specific genetic changes associated with differences in the pathogenicity of Marek's disease virus strains (GaHV-2) has been a formidable task due to the large number of mutations in mixed-genotype populations within DNA preparations. Very virulent UK isolate C12/130 induces extensive lymphoid atrophy, neurological manifestations and early mortality in young birds. We have recently reported the construction of several independent full-length bacterial artificial chromosome (BAC) clones of C12/130 capable of generating fully infectious viruses with significant differences in their pathogenicity profiles. Two of these clones (vC12/130-10 and vC12/130-15), which showed differences in virulence relative to each other and to the parental strain, had similar replication kinetics both in vitro and in vivo in spite of the fact that vC12/130-15 was attenuated. To investigate the possible reasons for this, the nucleotide sequences of both clones were determined. Sequence analysis of the two genomes identified mutations within eight genes. A single 494 bp insertion was identified within the genome of the virulent vC12/130-10 clone. Seven non-synonymous substitutions distinguished virulent vC12/130-10 from that of attenuated vC12/130-15. By sequencing regions of parental DNA that differed between the two BAC clones, we confirmed that C12/130 does contain these mutations in varying proportions. Since the individual reconstituted BAC clones were functionally attenuated in vivo and derived from a single DNA source of phenotypically very virulent C12/130, this suggests that the C12/130 virus population exists as a collection of mixed genotypes.


2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Qingdong Zeng ◽  
Fengping Yuan ◽  
Xin Xu ◽  
Xue Shi ◽  
Xiaojun Nie ◽  
...  

For map-based cloning of genes conferring important traits in the hexaploid wheat line 92R137, a bacterial artificial chromosome (BAC) library, including two sublibraries, was constructed using the genomic DNA of 92R137 digested with restriction enzymesHindIII andBamHI. The BAC library was composed of total 765,696 clones, of which 390,144 were from theHindIII digestion and 375,552 from theBamHI digestion. Through pulsed-field gel electrophoresis (PFGE) analysis of 453 clones randomly selected from theHindIII sublibrary and 573 clones from theBamHI sublibrary, the average insert sizes were estimated as 129 and 113 kb, respectively. Thus, theHindIII sublibrary was estimated to have a 3.01-fold coverage and theBamHI sublibrary a 2.53-fold coverage based on the estimated hexaploid wheat genome size of 16,700 Mb. The 765,696 clones were arrayed in 1,994 384-well plates. All clones were also arranged into plate pools and further arranged into 5-dimensional (5D) pools. The probability of identifying a clone corresponding to any wheat DNA sequence (such as geneYr26for stripe rust resistance) from the library was estimated to be more than 99.6%. Through polymerase chain reaction screening the 5D pools withXwe173, a marker tightly linked toYr26, six BAC clones were successfully obtained. These results demonstrate that the BAC library is a valuable genomic resource for positional cloning ofYr26and other genes of interest.


2007 ◽  
Vol 57 (1) ◽  
pp. 29-38 ◽  
Author(s):  
Daisuke Saisho ◽  
Eriko Myoraku ◽  
Shinji Kawasaki ◽  
Kazuhiro Sato ◽  
Kazuyoshi Takeda

2002 ◽  
Vol 61 (4) ◽  
pp. 441-446 ◽  
Author(s):  
David W. Burt ◽  
Paul M. Hocking

RésuméLa génomique de la volaille a bénéficié des avancements technologiques rapides acquis en génomique humaine et sur les organismes modèles. Certains outils et certains approches sont maintenant bien établis chez le poulet, y compris les cartes et marqueurs (génétiques comme physiques), mapping loci pour caractéres quantitatifs, mapping comparative, ressources expressed sequence tag et bacterial artificial chromosome, et mapping physique. De plus, la phase suivante de la découverte génétique, la génomique fonctionnelle, est en cours. Les progre`s dans le mapping de loci pour caracte`res quantitatifs de croissance et d'adiposité seront discutés pour illustrer ces nouvelles technologies et ces nouvelles approches dans l'étude de la génétique et de la physiologie avicole.


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