Cloning and sequencing of the gene Ahy-β encoding a subunit of peanut conarachin

• Published in: Plant science (Limerick) Vol. 168; no. 6; pp. 1387 - 1392
• Main Authors: Li, Hua-Guang, Wang, Lei, Zhang, Yi-Shun, Lin, Xiao-Dong, Liao, Bin, Yan, Yong-Sheng, Huang, Shang-Zhi
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 01.06.2005; Elsevier Science
• Subjects: 3' flanking region; amino acid sequences; Arachis hypogaea; Biological and medical sciences; CAAT box; complementary DNA; Conarachin; Fundamental and applied biological sciences. Psychology; gene expression regulation; Genes. Genome; legumin; Molecular and cellular biology; Molecular genetics; molecular sequence data; nucleotide sequences; oil crops; Peanut; peanuts; plant globulin-like protein; plant proteins; Promoter; promoter regions; Seed storage protein; sequence analysis; storage proteins; TATA box; tissue distribution; transcription factors; vicilin; Promoter; Seed storage protein; Conarachin; Peanut; Storage protein; Cotyledon; Seeds; Arachis hypogaea; Leguminosae; Complementary DNA; Gene; Dicotyledones; Aminoacid; Angiospermae; Spermatophyta
• ISSN: 0168-9452; 1873-2259
• DOI: 10.1016/j.plantsci.2004.09.028

A cDNA expression library of mid-maturation cotyledons of peanut ( Arachis hypogaea L. Shanyou 523) was constructed to clone the genes of peanut seed storage proteins. Six clones of cDNA were obtained by immunoscreening with a polyclonal antibody against a 65 kD subunit of conarachin from peanut seeds. The six cDNA clones could be divided into two groups, Ahy- α and Ahy-β. Furthermore, a complete cDNA sequence of Ahy- β was determined by using rapid amplification of cDNA ends (RACE). Northern blot analysis showed that Ahy- β was expressed only in developing peanut embryos. Sequence analysis revealed that the Ahy- β gene with 2507 bp was composed of four exons with 703, 176, 81 and 882 bp and three introns with 71, 72 and 73 bp. The deduced amino acid encoded a protein with 614 residues. Several well characterized elements for promoter were observed in the 342 bp promoter region of Ahy-β and included two TATA-boxes, four CAAT-boxes, a vicilin-box, a G-box, a RY-1 element, two RY-2 elements and a P-element, while a poly-A signal and two additional stop codons existed in the 3′UTR.