Four metallothionein (MT) isoforms have previously been identified in the brine shrimp Artemia. The goals of this study were to isolate at least one of the MT genes, clone a MT promoter region, and characterize the cis-acting sequences that regulate MT gene expression. PCR of Artemia genomic DNA amplified a product containing a MT gene with no internal introns. Three different approaches were made to clone the MT promoter. Firstly, conventional cloning of size-selected genomic DNA into a dephosphorylated vector produced empty clones or no clones. The use of a novel linear cloning vector produced many recombinant clones, but screening of the DNA library did not identify a MT promoter sequence. Lastly, genome walking by PCR amplified a sequence that lacked a TAT A box and metal response elements. A blast search identified the sequence as a mitochondrial hydroxyacylglutathione hydrolase found in marine organisms.
|Advisor:||Acey, Roger A.|
|Commitee:||Acey, Roger A., McAbee, Douglas D., Weers, Paulus M.|
|School:||California State University, Long Beach|
|Department:||Chemistry and Biochemistry|
|School Location:||United States -- California|
|Source:||MAI 51/04M(E), Masters Abstracts International|
|Subjects:||Molecular biology, Biochemistry|
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