Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Reexamination Certificate
2007-03-27
2007-03-27
Ketter, James (Department: 1636)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
C435S488000
Reexamination Certificate
active
10748055
ABSTRACT:
Provided is a method of producing a template DNA used for protein synthesis comprising a step of amplifying a linear double-stranded DNA by polymerase chain reaction (PCR), by using a reaction solution comprising a first double-stranded DNA fragment comprising a sequence coding for a protein or a portion thereof, a second double-stranded DNA fragment comprising a sequence overlapping with the 5′ terminal region of the first DNA fragment, a third double-stranded DNA fragment comprising a sequence overlapping with the 3′ terminal region of the first DNA fragment, a sense primer which anneals with the 5′ terminal region of the second DNA fragment, and an anti-sense primer which anneals with the 3′ terminal region of the third DNA fragment, wherein the second DNA fragment comprises a regulatory sequence for transcription and translation of a gene, and the concentrations of the second DNA fragment and the third DNA fragment in the reaction solution each range from 5 to 2,500 pmol/L. The use of this method enables efficient production of a template DNA for expression and purification of a protein.
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Kigawa Takanori
Motoda Yoko
Yabuki Takashi
Yokoyama Shigeyuki
Ketter James
Riken
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