کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
5905184 | 1159856 | 2016 | 6 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
A longer finger-subdomain of family A DNA polymerases found by metagenomic analysis strengthens DNA binding and primer extension abilities
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کلمات کلیدی
KBPEtdBrDNA polymerase from Thermus aquaticusethidium bromide - اتیدیوم برومایدAmino acids - اسید آمینه یا آمینو اسیدelectrophoretic mobility-shift assay - تحرک انتقال الکتریکی - تحرک تغییرkilobase pairs - جفت کیلوباEMSA یا electrophoretic mobility shift assay - سنجش تغییر تحرک الکتروفورتیکwild type - نوع وحشیpolymerase chain reaction - واکنش زنجیره ای پلیمرازPCR - واکنش زنجیرهٔ پلیمرازTaq polymerase - پلیکراز تاک
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
ژنتیک
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The family A DNA polymerases from thermophilic bacteria are useful for PCR. The DNA polymerase from Thermus aquaticus (Taq polymerase) was the original enzyme used when practical PCR was developed, and it has remained the standard enzyme for PCR to date. Knowledge gained from structure-function relationship studies of Taq polymerase is applicable to create PCR enzymes with enhanced performance. We collected the deduced amino acid sequences of the regions from motif A to motif C in the family A DNA polymerases from metagenomic sequence data, obtained by sequencing DNAs from microorganisms isolated from various hot spring areas in Japan. The corresponding regions of the polA gene for Taq polymerase were substituted with the metagenomic DNA gene fragments, and various chimeric DNA polymerases were prepared. Based on the properties of these chimeric enzymes and their sequences, we found an insertion sequence that affects the primer extension ability of the family A DNA polymerases. The insertion sequence is located in the finger subdomain, and it may enhance the affinity of the enzyme to DNA. Mutant Taq polymerases with the corresponding 9 amino acid insertion displayed enhanced PCR performance.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Gene - Volume 576, Issue 2, Part 1, 1 February 2016, Pages 690-695
Journal: Gene - Volume 576, Issue 2, Part 1, 1 February 2016, Pages 690-695
نویسندگان
Takeshi Yamagami, Hiroaki Matsukawa, Sae Tsunekawa, Yutaka Kawarabayasi, Sonoko Ishino, Yoshizumi Ishino,