High-efficiency T-vector cloning of PCR products by forced A tagging and post-ligation restriction enzyme digestion.
نویسندگان
چکیده
منابع مشابه
Construction of a Synthetic Vector for Preparation of a 100 Base Pair DNA Ladder
DNA size markers are widely used to estimate the size of DNA samples on agarose or polyacrylamide gelelectrophoresis (PAGE). DNA markers can be prepared by mixing PCR products with definite sizes.Alternatively, they are prepared by restriction enzyme digestion of the genomic DNA of bacteriophages ornatural and synthetic DNA plasmids. The present study describes engineering of ...
متن کاملطراحی و ساخت وکتور نوترکیب واجد ژن Vpr ویروس نقص ایمنی اکتسابی انسانی (HIV) با استفاده از وکتور EGFP-N1
Background and purpose: The purpose of this study was to design a recombinant vector pEFGP- N1 containing the full length of HIV-1Vpr gene. To the best of our knowledge, the cloning of Vpr gene in pEGFP_N1 is not previously done. Materials and methods: As a source of Vpr gene the pUC19-Vpr recombinant vector was confirmed by digestion with restriction enzymes BglII and NotI in order to separ...
متن کاملکلونینگ و توالییابی پلاسمید کد کننده پروتئین گرانولی متراکم 14 سویه RH توکسوپلاسما گوندیی
Background and purpose: Toxoplasmosis is a common parasitic disease throughout the world and one-third of the population has antibodies to Toxoplasma gondii. This disease causes serious medical problems in fetuses and immunocompromised individuals. As gene encoding protein GRA14 can be considered as a suitable target for DNA vaccine and designing diagnostic kits the aim of this study was to ...
متن کاملConstruction of a High Efficiency PCR Products Cloning T Vector Using pGEM-5zf (+)
A highly efficient cloning vector was constructed for cloning PCR products by inserting an 80 bp DNA fragment into pGEM-5zf (+) vector. The Xcm I digestion of this vector gave rise to a 3' overhanging deoxythymidine offering the possibility of cloning PCR products with 3' adenosine overhang created by Taq DNA polymerase. Furthermore, two EcoR I sites were added to the construct for identificati...
متن کاملCloning rhoptry protein 1 (ROP1) gene of Toxoplasma gondii (RH) in expression vector
Toxoplasma gondii contain various immunogenic antigens. The most important Toxoplasma antigens are somatic and excreted/secreted antigens. Rhoptry proteins are known as excreted/secreted antigens. These antigens have been proposed as a vaccine candidate against toxoplasmosis. The main objective of the present work was cloning rhoptry protein1 (ROP1) Gene of Toxoplasma gondii (RH) in a cloning...
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عنوان ژورنال:
- BioTechniques
دوره 23 5 شماره
صفحات -
تاریخ انتشار 1997