Method for synthesizing DNA sequentially

Chemistry: molecular biology and microbiology – Treatment of micro-organisms or enzymes with electrical or... – Modification of viruses

Patent

Rate now

  [ 0.00 ] – not rated yet Voters 0   Comments 0

Details

435 91, 435 68, 435820, 536 27, C12N 1500

Patent

active

042936527

ABSTRACT:
A method and a DNA linker are described for synthesizing relatively long double-stranded deoxyribonucleic acid sequences of defined composition. Short complementary single strand segments of oligonucleotides comprising part of the full sequence desired are synthesized using known procedures. Overlapping single strand segments are annealed forming double-stranded fragments which are inserted in cloning vectors and cloned in an appropriate host, both purifying the DNA fragments and amplifying the amount thereof. An adjacent fragment is then similarly synthesized in quantity and such fragments are inserted adjacent to the first synthetic introduced fragments in the cloning vectors, followed by cloning in an appropriate host. The procedure continues until the entire desired sequence has been formed, at which time it may be excised or cloned directly in the vectors upon which it was made. The described DNA linker contains a restriction enzyme recognition sequence for a restriction enzyme which cuts offset from the recognition sequence and exactly at the end of the linker, enabling synthesized DNA to be inserted at the end of the linker without disturbing the recognition sequence.

REFERENCES:
Itakura et al., Science, vol. 198, pp. 1056-1063 (1977).
Goebel et al., Genetic Engineering edited by Boyer et al., Elsevier/North-Holland Biomedical Press, pp. 47-58 (1978).
Greene et al., Methods in Molecular Biology, Chapter 4, pp. 87-111.
Hershfield et al., Proc. Natl. Acad. Sci., vol. 71, pp. 3455-3459 (1974).
Kleid et al., Proc. Natl. Acad. Sci., vol. 73, pp. 293-297 (1975).
Cohen et al., Proc. Natl. Acad. Sci., vol. 70, pp. 3240-3244 (1973).
Khorana et al., J. Mol. Biol., vol. 72, pp. 209-217 (1972).
Agarwal et al., Nature, vol. 227, pp. 27-34 (1970).
Khorana et al., J. Biol. Chem., vol. 251, pp. 565-570 (1976).
Carbon et al., Proc. Natl. Acad. Sci., vol. 72, pp. 1392-1396 (1975).
Polisky et al., Proc. Natl. Acad. Sci., vol. 73, pp. 3900-3902 (1976).
Maxam et al., Proc. Natl. Acad. Sci., vol. 74, pp. 560-564 (1977).
Sgaramella et al., J. Mol. Biol., vol. 72, pp. 427-444 (1972).
Itakura et al., J. Biol. Chem., vol. 250, pp. 4592-4597 (1975).

LandOfFree

Say what you really think

Search LandOfFree.com for the USA inventors and patents. Rate them and share your experience with other people.

Rating

Method for synthesizing DNA sequentially does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Method for synthesizing DNA sequentially, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method for synthesizing DNA sequentially will most certainly appreciate the feedback.

Rate now

     

Profile ID: LFUS-PAI-O-1207592

  Search
All data on this website is collected from public sources. Our data reflects the most accurate information available at the time of publication.