ISOLATION, CLONING AND CHARACTERIZATION OF ACTIN-ENCODING cDNAs FROM Jatropha curcas L. IP-2P

• Main Authors: Kinya Akashi, Akiho Yokota, Didy Sopandie, Utut Widyastuti1,2, Ratna Yuniati, Suharsono
• Format: Article
• Language: English
• Published: Universitas Indonesia 2011-11-01
• Series: Makara Seri Sains
• Subjects: actin; cDNA; cloning; gene isolation; Jatropha curcas L. IP-2P
• Online Access: http://journal.ui.ac.id/science/article/viewFile/1072/985

Actin is a major component of the plant cytoskeleton, so all cells contain this protein. Actin is expressed constitutivelyand is involved in basic housekeeping functions required for cell maintenance. Because of this, it has been frequentlyused as an internal control to normalize changes in gene expressions analysis. Actually, the information of nucleotidesequence of actin gene of Jatropha curcas L. population IP-2P from Indonesia is not available yet. The objective of thisresearch was to isolate, clone and characterize cDNA of actin genes of J. curcas IP-2P. Three partial actin genesequences had been successfully isolated by PCR using total cDNA as template, and actin primer designed fromconserved region of Arabidopsis thaliana. Nucleotide sequence analysis showed that the length of JcACT fragment is610, 534, and 701 bp encoding 203, 177, and 234 amino acids respectively. Local alignment analysis based on mRNAsequences shows that JcACT fragment shares 98% similarity with actin mRNA of Hevea brasiliensis and 99% withactin mRNA of Ricinus communis. Based on deduced amino acid sequence, JcACT is 100% identical to actins fromPrunus salicina, Gossypium hirsutum, and Betula luminifera. Even though these clones of cDNA are not completed yet,they can be used as reference in J. curcas L. gene expression analysis.