Date of Award

7-1-1999

Degree Type

Thesis

University or Center

Clark Atlanta University(CAU)

Degree Name

M.S.

Biological Sciences

First Advisor

Dr. John Browne

Abstract

Exon amplification is a method used to identify regions of DNA that contain transcribed sequences. The large cloning capacity of yeast artificial chromosome (YAC) systems and the inability to isolate large intact DNA from YACs introduce limitations to the exon amplification technique. The feasibility of exon amplification from a mega-YAC clone 2001C6 (CEPH) has been analyzed as a means to isolate transcribed sequences and has been used to produce 10 putative exon sequences from human chromosome 17q21. Six of the sequences showed homology to sequences that were previously published in the GenBANK database. Three of the sequences showed no homology, thereby indicating the isolation of putatively novel sequences. The sequences were radiolabeled and used as hybridization probes on a multiple tissue RNA dot blot. This blot contains RNA isolated from 50 human tissues. Clone E5 produced hybridization signals in fetal liver, fetal spleen and placenta. Clone F12 produced hybridization signals in fetal liver, fetal spleen, placenta and bone marrow. Clone G12 produced hybridization signals in all of the RNAs, indicating a pattern of expression similar to that of a housekeeping gene. These findings contribute to the enhancement of a high density transcriptional map within the q21-q22 region of human chromosome 17.

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