ISOLATION AND IDENTIFICATION OF A NEW PROTEIN FOR CAPPARIS SPINOSA RPS4 GENE | ||
| Biochemistry Letters | ||
| Article 7, Volume 8, Issue 1, 2012, Pages 165-199 PDF (770.46 K) | ||
| DOI: 10.21608/blj.2012.64268 | ||
| Authors | ||
| Faten Zahran1; Magdy M. Mohamed2; Mai Adel1 | ||
| 1Biochemistry Department, Faculty of Science, Zagazig University, Egypt | ||
| 2BicChemistry Department, Faculty of Science, Ain-Shams University. | ||
| Abstract | ||
| Capparis spinosa is arangland plant species growing in the desert of Egypt. It's one of the most important species in family cappariecia. Capparis spinosa is one of the medicinal plants established to have high diversity range of economic and medicinal value. Recently, the pharmacology and chemistry of this plant have been extensively studied. Chemical studies of the different parts of Capparis, both fermented and non-fermented, have shown the presence of many beneficial compounds. In this study the author selected a 606bp ribosomal protein 4s (rps4) chloroplast gene of Capparis spinosa, where the complete sequence of rps4 gene was amplified, sequenced, cloned, and expressed. rps4 gene is a gene in the land plant chloroplast genome and it is one of very popular marker in this genome. The forward primer started with the start codon ATG of rps4 and the reverse primer was designed to align aregion of 18nt upstream of the stop codon. The isolated gene is approximately 606 bp in lenght. After data alignment it was found that maximum identity was 98% with Capparis flexuosa ribosoma protein S4 (rps4) gene and 96% with Nasturtium officinale ribosomal protein S4 (rps4) gene. The information of this gene was found to be a new gene in comparison to the reported genes in the gene bank. According to the gene bank it's a new gene. Therefore it was given an accession number. The isolated gene was a complete gene and this gives a complete protein in expression where the accurate translation of the genetic message is an essential step in gene expretion. The clone was found to encode an 202 amino-acid protein fragment. It was expressed in Escherichia coli as a fusion protein of calculated molecular mass of about 48 kDa with Cterminus of glutathione-S-transferase ( about 26 kDa). The 202 amino-acid open reading frame encodes a protein with a calculated molecular mass of about 22 kDa. | ||
| Keywords | ||
| Capparis spinosa; rps4 gene; Sequencing; cloning; expression | ||
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