Cloning of mads regulatory gene from Theobroma cacao seeds

Roslina M.S., . and R. Kasran, . and Furtek D.B., . (2007) Cloning of mads regulatory gene from Theobroma cacao seeds. [Proceedings Paper]

There is currently an international effort in improving disease resistance and crop yield in Theobroma cacao L. one of an economically important crop of the Malaysia. We are developing an understanding in the molecular level of MADS box gene family encode transcription factors which play important roles in developmental control in plants animals. Degenerate PCR primers were designed to the highly conserved MADS DNA-binding domain and other conserved I intervenin and K keratin motifs. Other than these PCR primers preliminary screening with DNA and RNA was done with PCR primers from cocoa AGAMOUS TcAG. TcAG was in the cocoa flower expressed primarily in stamens and ovaries comparable to AG in Arabidopsis. Additionally study found that TcAG is also expressed in the fruit pod wall and during its entire development as well as in the fruit pulp. With the first aim of finding MADS box genes involved in the developing cocoa seeds we will isolate all members of the MADS-box gene family available and characterize them. Cloning of MADS genes from cocoa tissues is a necessary first step to investigate this gene at the molecular level. To accomplish this we have first designed MADS-specific primers for the isolation of the MADS gene expressed in seeds. mRNA was then purified from developing pods and first strand cDNA synthesized. The PCR primers were used to amplify partial dscDNA which were then cloned into plasmids and sequenced. Nucleotide and deduced amino acid sequences of the cDNA clones were compared and analyzed to other sequences in GenBank. A full-length MADS box gene sequences will provide enough information for further study such as examining their expression level between different stages in cocoa seed development.

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Abstract

There is currently an international effort in improving disease resistance and crop yield in Theobroma cacao L. one of an economically important crop of the Malaysia. We are developing an understanding in the molecular level of MADS box gene family encode transcription factors which play important roles in developmental control in plants animals. Degenerate PCR primers were designed to the highly conserved MADS DNA-binding domain and other conserved I intervenin and K keratin motifs. Other than these PCR primers preliminary screening with DNA and RNA was done with PCR primers from cocoa AGAMOUS TcAG. TcAG was in the cocoa flower expressed primarily in stamens and ovaries comparable to AG in Arabidopsis. Additionally study found that TcAG is also expressed in the fruit pod wall and during its entire development as well as in the fruit pulp. With the first aim of finding MADS box genes involved in the developing cocoa seeds we will isolate all members of the MADS-box gene family available and characterize them. Cloning of MADS genes from cocoa tissues is a necessary first step to investigate this gene at the molecular level. To accomplish this we have first designed MADS-specific primers for the isolation of the MADS gene expressed in seeds. mRNA was then purified from developing pods and first strand cDNA synthesized. The PCR primers were used to amplify partial dscDNA which were then cloned into plasmids and sequenced. Nucleotide and deduced amino acid sequences of the cDNA clones were compared and analyzed to other sequences in GenBank. A full-length MADS box gene sequences will provide enough information for further study such as examining their expression level between different stages in cocoa seed development.

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Item Type:	Proceedings Paper
Additional Information:	Summary En
AGROVOC Term:	THEOBROMA CACAO
AGROVOC Term:	SEEDS
AGROVOC Term:	CLONING
AGROVOC Term:	GENES
AGROVOC Term:	DNA
AGROVOC Term:	RNA
AGROVOC Term:	EXTRACTION
AGROVOC Term:	MALAYSIA
Geographical Term:	MALAYSIA
Depositing User:	Ms. Norfaezah Khomsan
Last Modified:	24 Apr 2025 05:27
URI:	http://webagris.upm.edu.my/id/eprint/15710

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