Magnetic bead capture를 이용한 염색체 띠 7q11, 7q22에 위치한 cDNA 선별

Abstract

[한국어 초록 없음]

Recently, surmountable amounts of genes are being cloned and it has become necessary to develop new techniques for discovering genes with known chromosomal location and their possible functions. We have developed one such a method and applied it to search for genes that may play a functional role during the development of the central nervous system. To observe the distribution of 18 weeks old fetal brain cDNAs on the chromosome, fluorescent in situ hybridization (FISH) was performed with biotin labeled 18 weeks old fetal brain cDNAs. Among the chromosome bands showing strong hybridization with the cDNAs, 7q11 and 7q22 were microdissected and amplified by PCR. Each band-specific probe pool was confirmed by FISH and band specific cDNAs were selected by magnetic bead capture method. Selected cDNAs were subcloned to plasmid vectors and the nucleic acid sequences were analysed. As a result 41 different clones from 7q11 and 43 different clones from 7q22 were obtained. They were categorized as 25 clones of well characterized genes, 22 clones showing low homology with known genes, 13 clones of simply registered uncharacterized human cDNAs, and 24 clones of unknown genes. From these results, it may be suggested that our technique is very useful to clone the genes expressed in the developing human brain with confirmed chromosomal location. In addition, this cloning technique can be used to discover the new genes related with neural development in combination with functional screening methods.