Human Recombinant DNA Library 



Larry L. Deaven, Robert K. Moyzis, Jon Longmire, and C. E. Hildebrand 



Life Sciences Division. Los Alamos National Laboratory, Los Alamos, NM 87345 



(505) 667-31 14, FTS 843-31 14 



The goal of the National Laboratory Gene Library Project (NLGLP) is the production 

 of chromosome-specific human gene libraries and their distribution to the scientific 

 community ( 1 ) for studies of the molecular biology of genes and chromosomes, 

 (2) for the study and diagnosis of genetic disease, and (3) for the physical mapping 

 (ordering) of chromosomes. This is a cooperative project employing the flow-sorting 

 and molecular-cloning expertise at the Los Alamos National Laboratory (LAND and 

 the Lawrence Livermore National Laboratory. The specific aim of Phase I of the project 

 was the production of complete digest libraries from each of the human chromosomal 

 types purified by flow sorting; the average insert size expected was about 4 kb. The 

 bacteriophage lambda vector was Charon 21 A, which has both EcoR I and Hind III 

 insertion sites accommodating human DNA fragments 0-9. 1 kb in size. Each laboratory 

 has produced a complete set of chromosome-specific libraries: LANL with EcoR I and 

 LLNL with Hind III. The small insert libraries are deposited in a repository at the 

 American Type Culture Collection, Rockville, Maryland: over 2000 aliquots have been 

 distributed to over 500 laboratories worldwide. 



The second phase of the project — the construction of partial digest libraries with larger 

 inserts in more advanced, recently developed lambda vectors (9-23 kb) and in cosmid 

 vectors (33—46 kb) — is under way. These large in.sert libraries have characteristics that 

 are better suited to basic studies of gene structure and function, organization of genes on 

 chromosomes, and ordering of cloned sequences. The Phase II strategy is to split the 

 genome between the two laboratories, with Livermore cloning 12 chromosomal types 

 (starting with 7, 1 1, 19, 21, 22, and Y) and Los Alamos cloning the other 12 (starting 

 with 4, 5, 8, 16. 17, and X). In this way, each chromosomal type will be cloned into 

 both lambda and cosmid vectors. Vectors currently in use include Charon 40 and 

 lambda GEMII (phage) and sCosl and Lawrist 5 (cosmid). Partial digest libraries 

 have been constructed in either phage or cosmid vectors for chromosomes X, Y, 16, 

 19, 21, and 22. 



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