کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5589786 | 1569831 | 2017 | 11 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Cloning and characterization of pyruvate carboxylase gene responsible for calcium malate overproduction in Penicillium viticola 152 and its expression analysis
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کلمات کلیدی
gDNArRNAPyCCSLBCCPTCARDNAITSRPMAMPSpecific primers - آغازگرهای خاصAmpicillin - آمپی سیلینribosomal deoxyribonucleic acid - اسید دز اکسید ریبونوکلئیک ریبوزومیribosomal ribonucleic acid - اسید ریبونوکلئیک ریبوزومیMalic acid - اسید مالیکBiotin - بیوتین، ویتامین B7،فاکتور W، کوآنزیم R، ویتامین HTaq - تاکbase pair - جفت پایهrevolutions per minute - سرعت در هر دقیقهMass spectrometry - طیف سنجی جرمیcorn steep liquor - مشروب تند ذرتunit - واحدbiotin carboxyl carrier protein - پروتئین حامل بیوتین کربوکسیلGenome walking - پیاده روی ژنومpyruvate carboxylase - پیرووات کربوکسیلازtricarboxylic acid cycle - چرخه اسید تریکاربوکیلیکkilobase - کیلوبایز
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
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چکیده انگلیسی
In this study, a pyruvate carboxylase gene (PYC) from a marine fungus Penicillium viticola 152 isolated from marine algae was cloned and characterized by using Genome Walking method. An open reading frame (ORF) of The PYC gene (accession number: KM593097) had 3582 bp encoding 1193 amino acid protein (isoelectric point: 5.01) with a calculated molecular weight of 131.2757 kDa. A putative promoter (intronless) of the gene was located at â 666 bp and contained a TATA box, several CAAT boxes, the 5â²-SYGGRG-3â² and a 5â²-HGATAR-3â² sequences. A consensus polyadenylation site (AATAAA) was also observed at + 10 bp downstream of the ORF. The protein deduced from the PYC gene had no signal peptide, was a homotetramer (4), and had the four functional domains. Furthermore, PYC protein also had three potential N-linked glycosylation sites, among them, -N-S-T-I- at 36 amino acid, -N-G-T-V- at 237 amino acid, and -N-G-S-S- at 517 amino acid were the most possible N-glycosylation sites. After expression of the PYC gene of P. viticola 152 in medium supplemented with CSL and biotin, it was found that the specific pyruvate carboxylase activity in MA production medium supplemented with CSL was much higher (0.5 U/mg) than in MA medium supplemented with biotin (0.3 U/mg), suggesting that optimal concentration of CSL is required for increased expression of the PYC gene, which is responsible for high level production of malic acid in P. viticola 152 strain.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 605, 20 March 2017, Pages 81-91
Journal: Gene - Volume 605, 20 March 2017, Pages 81-91
نویسندگان
Ibrar Khan, Sadia Qayyum, Shehzad Ahmed, Farhana Maqbool, Isfahan Tauseef, Kashif Syed Haleem, Zhen-Ming Chi,