[A novel T-cloning vector system].
The use of cloning vectors has revolutionized molecular biology. Any vector with appropriate cloning sites can be used to clone a section of DNA and polymerase chain reaction (PCR) is a useful method for producing DNA fragments that are intended to be cloned. When Taq polymerase is used in PCR for polymerization, the enzyme adds an extra adenosyl (A) nucleotide to the 3' end of the extended strand, in a template independent manner. T-cloning vectors are created by adding a thymidine (T) residue to the ends of the cloning site. This aids ligation with PCR products possessing A-T. In this study, we prepared a T-cloning vector system, which guarantees insertion of open reading frames to the right position for expression. This method provides an easy way of cloning PCR products assuring at the same time in frame insertion.
Köseoğlu
Ozgen
O
Hacettepe Universitesi Tip Fakültesi, Mikrobiyoloji ve Klinik Mikrobiyoloji Anabilim Dali, Ankara.
Kocagöz
Tanil
T
D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000737:chemistry / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics; D016133:Polymerase Chain Reaction; D013936:Thymidine / Q000737:chemistry / Q000235:genetics*
D004247:DNA; D015252:Deoxyribonucleases, Type II Site-Specific; C027440:GATATC-specific type II deoxyribonucleases; D013936:Thymidine