Identification of Mutants in Genes Encoding Arabidopsis Acyl-Coenzyme a Binding Proteins Acbp3, Acbp4 and Acbp5

This dissertation, "Identification of Mutants in Genes Encoding Arabidopsis Acyl-coenzyme A Binding Proteins ACBP3, ACBP4 and ACBP5" by Suk-wah, Chan, 陳淑華, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: Abstract of thesis entitled IDENTIFICATION OF MUTANTS IN GENES ENCODING ARABIDOPSIS ACYL-COENZYME A BINDING PROTEINS ACBP3, ACBP4 AND ACBP5 Submitted by CHAN Suk Wah for the Degree of Master of Philosophy at The University of Hong Kong in August 2004 Arabidopsis acyl-CoA binding proteins (ACBPs) that bind long-chain acyl-CoA esters can be divided into four classes: 1) cytosolic ACBP consisting of the acyl-CoA binding domain only; 2) membrane-associated ACBP1 and ACBP2 with ankyrin repeats; 3) ACBP3; and 4) kelch motif-containing ACBP4 and ACBP5. Cytosolic ACBPs, prevalent in eukaryotes, are involved in the storage and intracellular transport of long- chain acyl-CoAs. Except for cytosolic ACBPs, no other forms of ACBPs have been characterized in other eukaryotes. Arabidopsis cDNAs encoding membrane-associated ACBP1 and ACBP2, and those encoding ACBP3, ACBP4 and ACBP5 have been recently characterized. To elucidate the physiological functions of ACBP3, ACBP4 and ACBP5, an attempt was made to identify Arabidopsis. thaliana mutants in these genes using the reverse genetics approach. Putative ACBP3, ACBP4 and ACBP5 mutants were obtained from the Syngenta Arabidopsis Insertion Library (SAIL) and The Salk Institute Genome Analysis Laboratory (SALK) T-DNA insertional collections. The sites of T-DNA insertions in the putative ACBP3, ACBP4 and ACBP5 SAIL mutants were located upstream of their start codons; while putative ACBP3 and ACBP4 SALK mutants were located 3' of the respective translation start sites. Southern blot analyses showed the presence of T-DNA inserts in the SALK ACBP3 mutant, the SAIL ACBP4 mutant and the SAIL ACBP5 mutant. Reverse transcriptase-polymerase chain reaction (RT-PCR) analyses showed partial expression of ACBP3 mRNA, absence of ACBP4 mRNA and presence of ACBP5 mRNA expression in these mutants. Hence, only the SALK ACBP3 and ACBP4 mutants were deemed suitable for further analyses. DOI: 10.5353/th_b3621319 Subjects: Mutation (Biology) Acetylcoenzyme A Carrier proteins Arabidopsis thaliana - Genetics