amplification and cloning of taq dna polymerase gene from thermus aquaticus strain yt-1

نویسندگان

h. mir mohammad sadeghi

m. rabbani

f. moazen

چکیده

dna amplification using taq dna polymerase is one of the most widely used techniques in molecular biology and biotechnology. the aim of this study was to amplify the gene of this enzyme from a thermophilic bacteria called thermus aqauticus and clone it into a vector for future use. using specific primers the cdna of taq dna polymerase was amplified and ligated into the cloning vector ptz57r using ta cloning technique. the recombinant plasmids were identified using  restriction enzyme digestion. the presence of the taq dna polymerase gene was confirmed by dna sequencing. in conclusion, taq dna polymerase gene has been cloned in our laboratory and can be used for the production of large quantities of this enzyme.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Cloning and Expression of Thermus Aquaticus DNA Polymerase Gene, Using a Thermo-Inducible Expression Vector

DNA polymerase gene from Thermus aquaticus strain YT1 was amplified using VENTTM DNA po-lymerase and cloned under the control of X.PR promoter and expression was induced by a shift in tern perature. The culture was then sonicated, and after centrifugation the lysate was treated with poly‌ethyleneimine followed by a salting-out step. Finally the protein was precipitated with ammonium sulfate and...

متن کامل

cloning and expression of thermus aquaticus dna polymerase gene, using a thermo-inducible expression vector

dna polymerase gene from thermus aquaticus strain yt1 was amplified using venttm dna po-lymerase and cloned under the control of x.pr promoter and expression was induced by a shift in tern perature. the culture was then sonicated, and after centrifugation the lysate was treated with poly ethyleneimine followed by a salting-out step. finally the protein was precipitated with ammonium sulfate and...

متن کامل

Crystallization and preliminary X-ray diffraction studies of intact EF-Tu from Thermus aquaticus YT-1.

Many attempts have been made to elucidate the three-dimensional structure from elongation factor Tu, but so far the only crystals suitable for X-ray crystallography contained a partially degraded protein. Here, we report the crystallization of a fully active, intact EF-Tu from thermus aquaticus. The crystals belong to hexagonal space group P6(3)(22) and diffract up to 2.6 A. The cell dimensions...

متن کامل

Gene walking by PCR amplification of short fragments from Taq DNA polymerase--modified P1 plasmid DNA and TA cloning.

1.Belgrader, P., W. Benett, D. Hadley, G. Long, R. Mariella, Jr., F. Milanovich, S. Nasarabadi, W. Nelson et al. 1998. Rapid pathogen detection using a microchip PCR array instrument. Clin. Chem. 44:2191-2194. 2.Belgrader, P., J.K. Smith, V.W. Weedn and M.A. Northrup. 1998. Rapid PCR identity testing using a battery-powered miniature thermal cycler. J. Forensic Sci. 43:315-319. 3.Belinda, A.J.G...

متن کامل

Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus.

The thermostable properties of the DNA polymerase activity from Thermus aquaticus (Taq) have contributed greatly to the yield, specificity, automation, and utility of the polymerase chain reaction method for amplifying DNA. We report the cloning and expression of Taq DNA polymerase in Escherichia coli. From a lambda gt11:Taq library we identified a Taq DNA fragment encoding an epitope of Taq DN...

متن کامل

High fidelity DNA synthesis by the Thermus aquaticus DNA polymerase

We demonstrate that despite lacking a 3'----5' proofreading exonuclease, the Thermus aquaticus (Taq) DNA polymerase can catalyze highly accurate DNA synthesis in vitro. Under defined reaction conditions, the error rate per nucleotide polymerized at 70 degrees C can be as low as 10(-5) for base substitution errors and 10(-6) for frameshift errors. The frequency of mutations produced during a sin...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
research in pharmaceutical sciences

جلد ۱، شماره ۱، صفحات ۴۹-۰

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023