Characterization of β-Tubulin cDNA(s) from Catharanthus roseus
Journal Title: Journal of Biodiversity and Environmental Sciences (JBES) - Year 2015, Vol 6, Issue 1
Abstract
Vinblastine and vincristine are anti-tumor drugs produced by different species of Vinca. Vinca alkaloids depolymerize microtubules and as a result interfere with the normal formation of microtubules during cell division. Extremely low amount of vinca alkaloids in Vinca species and increased demand for chemotherapy are motives to scale up its production by biotechnological approaches. Here, β-tubulin gene family in Catharanthus roseus (vinca rosea) was analyzed to provide enough information for further approaches. 24 β-tubulin coding sequences from plants, mainly Arabidopsis thaliana, were obtained from genebank and aligned to design gene specific primers. Total RNA from leaves of C. roseus were isolated, the cDNA(s) synthesized using specific reverse primers, the amplified fragments cloned in pJET cloning vector and sequenced. Eight clones containing partial β-tubulin cDNAs’ of C. roseus; including an almost full-length coding sequence, were constructed. The comparison of sequencing results with β-tubulin mRNA from Arabidopsis thaliana, Arabidopsis lyrata, Populus trichocarpa, Zea maize, Oryza sativa, Hordeum vulgare, Gossypium hirsutum, Nicotiana attenuate, Solanum tuberosum and lycopersicone sculentum revealed about 85% similarity of nucleotide sequence and about 98% identities at amino acid level. The clustalW alignment analyses underlined a short peptide at position 359-367 with substitutions very specific to C. roseus. Based on our sequencing results, we predicted at least four members of β-tubulin gene family for C. roseus. TUB cDNAs’ from C. roseus sequenced to reveal any differentiation in comparison with non-vinca alkaloid producer plants. It was proposed the sites would carry higher potential for any amino acid alteration providing self-protection in vinca alkaloids producer plants; e.g. C. roseus.
Authors and Affiliations
Ziba Fooladvand, Shamsozoha Abolmaali, Abbas Saidi, Hamed Ashourion
Keywords
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