Construction and Characterization of Large-insert Genomic Libraries (BAC and Fosmid) from the Ascidian Botryllus Schlosseri and Initial Physical Mapping of a Histocompatibility Locus

Overview

Journal Mar Biotechnol (NY)

Specialties Biology
Biotechnology

Date 2003 Jul 24

PMID 12876644

Citations 12

Authors

Anthony W De Tomaso

Irving L Weissman

Affiliations

Soon will be listed here.

Abstract

The colonial protochordate Botryllus schlosseri is genetically manipulable and represents a potential model organism for a variety of biological disciplines, including immunology, stem cell biology and development. This article presents the construction and characterization of both BAC and fosmid genomic libraries of the 725-Mbp B. schlosseri genome. The BAC library currently consists of 2x genome coverage with an average insert size of 80 kb. The fosmid library is at 11x genome coverage with an average insert of 40 kb. B. schlosseri is a small organism containing a large number of compounds that hinder DNA purification. Thus a number of protocols had to be modified in order to make purified, high molecular weight inserts for cloning, including both gel purification and insert concentration techniques. Both libraries were characterized by using them in initial physical mapping of a single histocompatibility locus, and were found to be representative and functional. These libraries are important tools for physical mapping and positional cloning in the B. schlosseri genome, and the techniques adapted to make them are suitable for use on other organisms in which high molecular weight DNA is difficult to purify.

Citing Articles

Establishment of an Efficient and Flexible Genetic Manipulation Platform Based on a Fosmid Library for Rapid Generation of Recombinant Pseudorabies Virus.

Zhou M, Abid M, Yin H, Wu H, Teklue T, Qiu H Front Microbiol. 2018; 9:2132.

PMID: 30233561 PMC: 6133995. DOI: 10.3389/fmicb.2018.02132.

Allorecognition proteins in an invertebrate exhibit homophilic interactions.

Karadge U, Gosto M, Nicotra M Curr Biol. 2015; 25(21):2845-2850.

PMID: 26455308 PMC: 4631698. DOI: 10.1016/j.cub.2015.09.030.

Molecular evolution and in vitro characterization of Botryllus histocompatibility factor.

Taketa D, Nydam M, Langenbacher A, Rodriguez D, Sanders E, De Tomaso A Immunogenetics. 2015; 67(10):605-23.

PMID: 26359175 PMC: 11614195. DOI: 10.1007/s00251-015-0870-1.

Botryllus schlosseri allorecognition: tackling the enigma.

Taketa D, De Tomaso A Dev Comp Immunol. 2014; 48(1):254-65.

PMID: 24709050 PMC: 4185259. DOI: 10.1016/j.dci.2014.03.014.

Advances in BAC-based physical mapping and map integration strategies in plants.

Ariyadasa R, Stein N J Biomed Biotechnol. 2012; 2012:184854.

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