We claim:
1. A nucleic acid with a 5 end and a 3 end comprising a first functional nucleotide sequence and a scissile strand topoisomerase I cleavage motif sequence, wherein the scissile strand topoisomerase I cleavage motif sequence is located 3 to the first functional nucleotide sequence and provides a scissile strand topoisomerase I cleavage site that is not more than 10 bases from the 3 end of the nucleic acid.
2. The nucleic acid of claim 1, wherein the scissile strand topoisomerase cleavage motif sequence is selected from the group consisting of: CCCTT and TCCTT.
3. The nucleic acid of claim 1, wherein the first functional nucleotide sequence is selected from the group consisting of: a prokaryotic promoter sequence, a eukaryotic promoter sequence, a viral promoter sequence, a mutational sequence, a polypeptide tag encoding sequence, a nucleic acid tag sequence, a terminator sequence, a fusible protein encoding sequence, a radioactively labeled nucleotide sequence, a chemically labeled nucleotide sequence and an intronic sequence.
4. An adaptor comprising a first nucleic acid with a 5 end and a 3 end and comprising a scissile strand topoisomerase I cleavage motif having a 5 motif sequence contiguous with a 3 motif terminal nucleotide, said 3 motif terminal nucleotide being contiguous with a palindromic sequence of not less than two nucleotides nor more than 10 nucleotides and said palindromic sequence being contiguous with a 3 end nucleotide that is complementary to the 3 motif terminal nucleotide of the scissile strand topoisomerase I cleavage motif.
5. The adaptor of claim 4 further comprising a second nucleic acid having a 5 end sequence that is complementary to the 5 motif sequence of the scissile strand topoisomerase I cleavage motif.
6. The first nucleic acid of the adaptor of claim 4, wherein the 3 motif terminal nucleotide of the scissile strand topoisomerase I cleavage motif is T and the 5 motif sequence of the scissile strand topoisomerase I cleavage motif is selected from the group consisting of CCCT and TCCT.
7. The first nucleic acid of the adaptor of claim 4 further comprising a restriction endonuclease site located 5 to the scissile strand topoisomerase I cleavage motif.
8. The first nucleic acid of the adaptor of claim 4 further comprising a 5 end sequence that is complementary to the 5-overhang of a restriction endonuclease site.
9. The first nucleic acid of claim 7 or claim 8, wherein the restriction endonuclease is selected from the group consisting of: BamH I, Bgl II, Cla I, Dde I, Eae I, Eag I, EcoR I, Hind III, Kas I, Mbo I, Mlu I, Nco I, Nde I, Nhe I, Not I, PaeR7 I, Sal I, Sau3A, Spe I, Sty I, Xba I, Xho I and Xma I.
10. The first nucleic acid of the adaptor of claim 4, further comprising a first functional nucleotide sequence selected from the group consisting of: a prokaryotic promoter sequence, a eukaryotic promoter sequence, a viral promoter sequence, a mutational sequence, a polypeptide tag encoding sequence, a nucleic acid tag sequence, a terminator sequence, a fusible protein encoding sequence, a radioactively labeled nucleotide sequence, a chemically labeled nucleotide sequence and an intronic sequence.
11. A method for joining an adaptor sequence to a target nucleic acid sequence comprising:
providing a nucleic acid adaptor of claim 5,
providing a target nucleic acid with a one base 3 overhang nucleotide that is complementary to the 3 motif terminal nucleotide of the scissile strand topoisomerase cleavage motif, and
incubating the nucleic acid adaptor with the target nucleic acid in the presence of a topoisomerase I activity,
thereby joining the adaptor sequence to the target nucleic acid sequence.
12. The method of claim 11, wherein the first nucleic acid of the adaptor of claim 5 further comprises a functional nucleotide sequence that is 5 to the scissile strand topoisomerase I cleavage motif.
13. The method of claim 12, wherein the functional nucleotide sequence is selected from the group consisting of: a prokaryotic promoter sequence, a eukaryotic promoter sequence, a viral promoter sequence, a mutational sequence, a polypeptide tag encoding sequence, a nucleic acid tag sequence, a terminator sequence, a fusible protein encoding sequence, a radioactively labeled nucleotide sequence, an intronic sequence.
14. The method of claim 12, wherein the functional nucleotide sequence is a phage promoter selected from the group consisting of: an SP6 promoter, a T3 promoter and a T7 promoter.
15. The method of claim 11, further comprising the step of amplifying the joined product.
16. The method of claim 15, wherein the joined product is amplified by a polymerase chain reaction utilizing a first primer specific to the nucleic acid adaptor and a second primer specific to the target nucleic acid sequence.
17. The method of claim 11, wherein the target nucleic acid is generated by a polymerase chain reaction of a target genomic or a target cDNA sequence with a 5 sense strand primer and a 3 anti-sense strand primer.
18. The method of claim 17, wherein the adaptor provides a functional nucleotide sequence that is a promoter sequence and further comprising the steps of preparing at least two separate amplification reactions from the joined product comprising:
a first amplification reaction with 3 anti-sense strand primer and a first adaptor primer; and
a second amplification reaction with a 5 sense strand primer and a second adaptor primer, wherein the first adaptor primer comprises a sequence in the first nucleic acid of the adaptor and the second adaptor primer comprises a sequence in the second nucleic acid of the adaptor.
19. The method of claim 18 further comprising the step of isolating the product of either the first amplification reaction or the second amplification reaction.
20. The method of claim 19 further comprising contacting the amplification product with an RNA polymerase activity which recognizes said promoter sequence.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
What is claimed is:
1. An intubating laryngoscope system providing for ready interchange of blades, for video display, and for convenience in use comprising:
a handle to be held by an anesthesiologist;
a blade having a surface configured to enable the anesthesiologist to elevate the tongue of a patient, a proximal end, and a distal end;
a hinge-like joinder separably to join said handle and said blade, said joinder having a first portion on said handle and a second portion on said proximal end of said blade, one of said portions having a rod and the other portion having a hook, whereby in an engaged orientation the hook and the rod are engaged, and in a released configuration they may be separated, the handle and blade being rigidly attached to one another in the engaged orientation;
a flexible cable extending from said handle having a fixed end fixed to the handle, and a free end;
a guide tube attached to said blade, extending from near the proximal end of the blade toward its distal end, said guide tube including an enlarged a funnel-like entry portion to facilitate the entry of said flexible cable into said guide tube, the length of said flexible cable being such that when the handle and blade are in the engaged orientation, the flexible cable is disposed in a predetermined position in the blade;
a camera;
a light source;
a light-transmitting fiber optic in said flexible cable;
image-transmitting means in said flexible cable;
a second cable extending from said handle;
a video receiver connected to said second cable receptive to signals from said camera; and
a video screen receptive to said video receiver to display images transmitted from said camera.
2. A system according to claim 1 in which said camera is disposed in a chamber in a said standard handle, and said image-transmitting flexible cable comprises a coherent fiber optic bundle having one end exposed to a field of view and a second end exposed to said camera.
3. A system according to claim 1 in which said light source is separate from the handle, and in which said light-transmitting fiber optic is adapted to receive light from said light source to be transmitted to the field of view.
4. A system according to claim 3 in which said camera is disposed in a chamber in said standard handle, and said flexible cable includes a coherent fiber optic bundle having one end exposed to a field of view and a second end exposed to said camera.
5. A system according to claim 1 in which said camera is mounted to the distal end of the flexible cable, and the image-transmitting means is a lead which conveys image data through said handle to said video receiver.
6. A system according to claim 1 in which said light source is a battery and bulb combination in said handle to provide light to said light-transmitting means.
7. An intubating laryngoscope system enabling relay interchange of blades, video display, and convenient use, comprising:
a handle to be held by an anesthesiologist;
a blade having a surface configured to enable the anesthesiologist to elevate the tongue of a patient, a proximal end, and a distal end;
a hinge-like joinder separably to join said handle and said blade, said joinder having a first portion on said handle and a second portion on said proximal end of said blade, one of said portions having a rod and the other portion having a hook, whereby in an engaged orientation the hook and the rod are engaged, and in a released configuration they may be separated, the handle and blade being rigidly attached to one another in the engaged orientation;
a flexible cable extending from said handle having a fixed end fixed to the handle, and a free end;
a guide tube attached to said blade, extending from near the proximal end of the blade toward its distal end, said guide tube including an enlarged funnel-like entry portion to facilitate the entry of said flexible cable into said guide tube, the length of said flexible cable being such that when the handle and blade are in the engaged orientation, the flexible cable is disposed in a pre-determined position in the blade;
a camera;
a light-transmitting fiber optic in said flexible cable;
image-transmitting means in said flexible cable; and
a second cable extending from said handle; and
said second cable being adapted to be connected to a video receiver which is receptive to signals from said camera.
8. A laryngoscope according to claim 7 in which said camera is disposed in a chamber in said standard handle, and said image-transmitting flexible cable comprises a coherent fiber optic bundle having one end exposed to a field of view and a second end exposed to said camera.
9. A laryngoscope according to claim 7 in which said light transmitting fiber optic in said flexible cable is adapted to receive light from a separate light source to be transmitted to the field of view.
10. A laryngoscope according to claim 9 in which said camera is disposed in a chamber in said standard handle, and said flexible cable includes a coherent fiber optic bundle having one end exposed to a field of view and a second end exposed to said camera.
11. A laryngoscope according to claim 7 in which said camera is mounted to the distal end of the flexible cable, and the image-transmitting means is a lead from said camera extending through said handle for conveying image data through said handle to a video receiver.
12. A laryngoscope according to claim 7 in which said light source is a battery and bulb combination in said handle, to provide light to said light-transmitting means.
13. A system according to claim 1 in which said guide tube has a closed distal end with an optically clear and transparent window at said distal end, and a wall extending to said entry portion, said wall and said window being imperforate and sterilizeable, whereby a non-sterile and flexible cable can be inserted into said guide tube, and the guide tube disposed of after use without need to sterilize the flexible cable before its next use.
14. A laryngoscope according to claim 7 in which said guide tube has a closed distal end with an optically clear and transparent window at said distal end, and a wall extending to said entry portion, said wall and said window being imperforate and sterilizeable, whereby a non-sterile said flexible cable can be inserted into said guide tube, and the guide tube disposed of after use without need to sterilize the flexible cable before its next use.
15. A system according to claim 1 in which a device to remove the geometric pattern from the signal to the video screen receives and processes the image signal form the camera.
16. A laryngoscope system according to claim 1 in which a conduit mounted to the blade with an exit opening near the distal end of the flexible cable is adapted to discharge a stream of oxygen adjacent to the distal end of the guide tube.
17. A laryngoscope according to claim 7 in which a conduit mounted to the blade with an exit opening near the distal end of the flexible cable is adapted to discharge a stream of oxygen adjacent to the distal end of the guide tube.