Our efforts in 1992 have been focused on expanding this library, characterizing specific clones isolated from the library, and demonstrating the use of BACs and Fosmids in creating physical maps. Our goal is to create a 4-5X library which will be accessible for screening both by colony hybridization and by PCR. In January 1992 we began construction of an arrayed total human genomic library prepared in our BAC vector. which will permit detailed analysis of complex genomes. Over the past several years we have been developing a new approach to cloning large fragments of mammalian DNA in E. We will further optimize the technology so that libraries of fragments larger than 200 kb can be readily prepared. We have thus far prepared two human chromosome 22 specific Fosmid libraries and we are in the process of constructing a chromosome 22 specific BAC library composed of fragments larger than 100 kb. We, therefore, plan to use the large fragment cloning procedure as well as a variety of other more » approaches to generate a complete map of overlapping clones corresponding to human chromosome 22. The ultimate test of the effectiveness of cloning and mapping technology is the degree to which it can be efficiently applied to solve complex mapping problems. The cosmid size clones are called Fosmids and the clones containing larger inserts (100-200 kb) are called bacterial artificial chromosomes (BACs).
In addition to large size, these systems are capable of maintaining human DNA with a high degree of stability. We have developed bacterial F-factor based systems for cloning large fragments of human DNA in E. We will further optimize the technology so that libraries of fragments larger than 200 kb can be readily = , We, therefore, plan to use the large fragment cloning procedure as well as a variety of other approaches to generate a complete map of overlapping clones corresponding to human chromosome 22.
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