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 Title: |
US4407948:
Purification of nucleotide sequences suitable for expression in bacteria
[ Derwent Title ]
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| Country: |
US United States of America
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| Inventor: |
Goodman, Howard M.; San Francisco, CA
Shine, John; San Francisco, CA
Seeburg, Peter H.; San Francisco, CA
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| Assignee: |
The Regents of the University of California, Berkeley, CA
other patents from UNIVERSITY OF CALIFORNIA, THE REGENTS OF (599425) (approx. 4,840)
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| Published / Filed: |
1983-10-04
/ 1982-02-05
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| Application Number: |
US1982000346123
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| IPC Code: |
Advanced:
C07K 14/575;
C07K 14/61;
C07K 14/62;
C12N 15/10;
IPC-7:
C12N 15/00;
C12P 19/34;
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| ECLA Code: |
C07K14/575D; C07K14/61; C07K14/62; C12N15/10D;
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| U.S. Class: |
Current:
435/091.52;
435/091.53;
435/270;
536/023.1;
930/010;
930/120;
Original:
435/091;
435/172;
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| Field of Search: |
435/093,91,172
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| Government Interest: |
The Government has rights in this invention pursuant to Grants No. GM-21830 and CA 14026 awarded by the Dept. of Health, Education and Welfare.
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| Priority Number: |
| 1982-02-05 |
US1982000346123 |
| 1977-09-23 |
US1977000836218 |
| 1978-04-19 |
US1978000897710 |
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| Abstract: |
A method for purifying a DNA fragment of specific desired nucleotide sequence containing a restriction site, from a population of DNA molecules homogeneous in length by endonuclease cleavage and fractionation.
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| Attorney, Agent or Firm: |
Keil & Witherspoon ;
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| Primary / Asst. Examiners: |
Tanenholtz, Alvin E.;
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| INPADOC Legal Status: |
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| Parent Case: |
This is a division of application Ser. No. 897,710 filed Apr. 19, 1978, now U.S. Pat. No. 4,363,877, and a continuation-in-part of copending application, Ser. No. 836,218, filed Sept. 23, 1977 and now abandoned.
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| Family: |
Show 157 known family members
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First Claim:
Show all 9 claims |
What is claimed is:
1. A method of purifying linear DNA of specific nucleotide sequence containing a restriction site, from an essentially homogeneous length population of DNA molecules, comprising the steps of:
- (a) pretreating the DNA to remove any 5' phosphate end groups,
- (b) incubating the DNA with a restriction endonuclease capable of acting on the specific desired nucleotide sequence to produce two linear sub-fragments thereof,
- (c) fractionating the sub-fragments according to their length,
- (d) isolating the two sub-fragments,
- (e) rejoining the sub-fragments covalently, in a DNA ligase catalyzed reaction, to reconstitute the original specific sequence, and
- (f) fractionating the rejoined DNA molecules according to their length, whereby the specific sequence is purified from a population of DNA molecules of essentially homogeneous length.
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| Background / Summary: |
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| Drawing Descriptions: |
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| Description: |
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| Forward References: |
Show 22 U.S. patent(s) that reference this one
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