I think this fits here??... 164.195.100.11
United States Patent
6,054,299
Conrad
April 25, 2000
Stem-loop cloning vector and method
Abstract
Methods and compositions for the isolation and specific targeting of any single stranded DNA sequence to be acted upon by
any desired double stranded DNA genetic element or recognition sequence in a cis-oriented fashion. The invention involves
construction of a vector comprised of single stranded sequences that form "stem-loop" structures wherein the "loop" comprises
the single stranded target sequence and the "stem" comprises the double stranded "functional" cis-acting genetic elements or
recognition sequences. The in vivo formation of this single stranded intermediate (prior to stem-loop folding) is performed by
genetic elements which direct the normal DNA replication of any one of a number of prokaryotic and eukaryotic viruses during
their life cycles. Constructs containing these replicative functions are used to produce the desired single stranded intermediates.
Also included in the stem-loop structure is an appropriate inverted tandem repeat which, forms the "stem" of the stem-loop
structure. Once these functions are designed into a particular chimera, any desired functional genetic elements (i.e., promoters,
enhancer elements, or ribosome binding sites) or recognition sequences (i.e. restriction endonuclease sites) may comprise the
inverted repeats and thus the cis-acting double stranded "stem" directs these useful functions to the isolated single stranded
target sequence in the "loop".
Inventors:
Conrad; Charles A. (5807 Miller Valley, Houston, TX 77066)
Appl. No.:
877251
Filed:
June 17, 1997
U.S. Class:
435/91.1; 435/91.4; 435/320.1; 435/252.3
Intern'l Class:
C12N 015/00
Field of Search:
435/172.3,320.1,91.1,252.3
References Cited [Referenced By]
U.S. Patent Documents
Other References
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Primary Examiner: Martinell; James
Attorney, Agent or Firm: Wisner; Mark R. |
