Open Access

ARTICLE

Cloning and characterization of 66 kDa streptavidin-binding peptides (SBP) of Pisum sativum L. embryo specific to var. Alaska

Mahmoud MOUSTAFA^1,2 , Saad ALAMRI¹, Tarek TAHA³, Ali SHATI¹, Sulaiman ALRUMMAN¹, Mohamed ALKAHTANI¹

1 Department of Biology, College of Science, King Khalid University, 9004, Abha, Kingdom of Saudi Arabia
2 Department of Botany, Faculty of Science, South Valley University, Qena, Egypt
3 Environmental Biotechnology Department, Genetic Engineering and Biotechnology Research Institute (GEBRI), City of Scientific Research and Technological Applications (SRTA-City), New Borg El-Arab City, 21934, Alexandria, Egypt

* Address correspondence to: Mahmoud Moustafa,

BIOCELL 2019, 43(3), 155-166. https://doi.org/10.32604/biocell.2019.06814

Abstract

The aim of the current research was to clone and to characterize the partial 66 kDa streptavidin-binding peptide (SBP) found in the germinated embryos of Pisum sativum L. var. Alaska. The pea (P. sativum var. Alaska) embryos possess prominent 66 kDa SBPs that gradually disappeared after few hours of germination in germinated embryos, but not in the cotyledons. The total RNA was isolated from embryos of P. sativum but could not be isolated from the cotyledons. The partial nucleotides sequences of 66 kDa SBPs of embryonic stalk (P. sativum var. Alaska) were cloned and identified using pMOSBlue vector. 66 kDa (SBP) gene from the embryos of P. sativum var. Alaska possesses 327 bp having an open reading frame (ORF) region in a part of the gene that encoded for 108 amino acids. Alignment showed similarity among 66 kDa SBPs P. sativum var. Alaska, with P. sativum seed biotinylated protein (SBP65) and P. sativum sbp65a mRNA with DNA distance matrix between 0.0094 to 1.2676. MALDI-TOF mass spectrometry analysis of 66 kDa (SBP) proteins showed it had similar short peptides to 19 proteins found in different organisms, especially Convicilin precursor, and the seed biotinylated protein in P. sativum. The alignment results of both nucleotide sequences and amino acid residues either from cloning or MALDI-TOF-MS showed differences with related species, especially P. sativum. No mRNA was found in the cotyledons during seeds germination, which means no metabolic activities and this part may act only as food reservoirs for growing newly embryos.

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