1. A resistance device for an exerciser with an external design thereof in a unitary machine; application of the present invention to an exerciser minimizes the volume thereof and obtains the effect of saving the most cost; the structure of the present invention mainly comprises external cases of left and right machines, a driving resistance disk, a positioning ring, a adjustable resistance disk, an oblique Y-shaped clamp, a tension spring, a main shaft and a drawing steel rope; wherein:
the external cases of left and right machines are locked in clamping type and filled with machine oil of high viscosity and a main shaft positioned and sealed through the cooperation of a shaft bearing and a oil seal to allow the driving resistance disk, the positioning ring, the adjustable resistance disk, the oblique Y-shaped clamp and the tension spring to exist and operate therein;
the driving resistance disk has a unitary belt wheel penetrated by the main shaft; the internal and external portions of the wheel are disposed with the shaft bearings and the oil seals; it is capable of making spinning rotation and forming a corresponding structure with the adjustable resistance disk; the front plane of the disk is disposed with a plurality of concentric circular pieces staggered and inserted with resistance pieces of the adjustable resistance disk; a proper gap is reserved between the resistance pieces; the innerest rims of the resistance pieces are disposed with circular oil lead holes and the resistance pieces are disposed with inclined and convergent slot ways; the backside of the disk is disposed with diagonal slot ways for rapidly leading the machine oil into all the resistance pieces;
the adjustable resistance disk is penetrated by the main shaft and controlled by the positioning ring, the oblique Y-shaped clamp and the tension spring; it is not capable of making spinning rotation; the front aspect of the disk is stopped and positioned by the positioning ring; the rear aspect thereof is forced by the tension spring to form a corresponding structure with the driving resistance disk; the front plane of the disk is disposed with a plurality of concentric circular pieces staggered and inserted with resistance pieces of the driving resistance disk; a proper gap is reserved between the resistance pieces; the innerest rims of the resistance pieces are disposed with circular oil lead holes and the resistance pieces are disposed with inclined and convergent slot ways; the backside of the disk is disposed with diagonal slot ways for rapidly leading the machine oil into all the resistance pieces;
the oblique Y-shaped clamp is inclined and slightly resilient; the hollow section at the bottom portion thereof straddles on the main shaft with two end foot portions respectively fastened on the backside of the adjustable resistance disk; the central position on the oblique Y-shaped clamp is fastened with the drawing steel rope penetrated into the external case of the machine; the top end point of the said oblique Y-shaped clamp is fixed onto the inner rim wall of the external case of the machine to draw the adjustable resistance disk via the drawing steel rope to form a micro adjusting function of displacing forwardly and backwardly.
2. A resistance device for an exerciser according to claim 1, wherein the oil lead slot ways disposed on the driving resistance disk and the adjustable resistance disk are not of a corresponding structure.
3. A resistance device for an exerciser according to claim 1, wherein the adjustable resistance disk can be a member of two semi-spheres covered externally onto the outer rim side of the driving resistance disk; the inner and outer rims of the resistance pieces of two disks are correspondingly staggered and inserted to enable the driving resistance disk to make spinning rotation under the applied external force and the adjustable resistance disk to have function of expanding displacement so as to achieve the equivalent effect of forming resistance and adjusting resistance value.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1-20. (Canceled)
21. A method of producing a transgenic mouse comprising a vector, comprising
a) introducing a vector into murine embryonic stem (ES) cells, wherein the vector comprises:
i) a 5 gene trap cassette, comprising in operable combination:
1) a splice acceptor;
2) a first exon sequence located 3 to said splice acceptor, said first exon sequence encoding a marker enabling the identification of a cell expressing said first exon sequence; and
3) a polyadenylation sequence located at the 3 end of said first exon sequence;
ii) a 3 gene trap cassette located 3 to said polyadenylation sequence, comprising in operable combination:
1) a first promoter;
2) a second exon sequence located 3 from and expressed by said first promoter, said second exon sequence not encoding an activity conferring antibiotic resistance;
3) a splice donor sequence located at the 3 end of said second exon sequence; and
wherein said vector does not encode a promoter mediating the expression of said first exon sequence, and wherein said vector does not encode a sequence that mediates the polyadenylation of an mRNA transcript encoded by said second exon sequence;
b) selecting a murine ES cell that comprises the vector; and
c) making a transgenic mouse comprising the vector from the selected murine ES cell that comprises the vector.
22. The method of claim 21, wherein the vector from the selected murine ES cell that comprises the vector is non-homologously incorporated into the genome of at least one cell in the transgenic mouse.
23. The method of claim 22, further comprising identifying at least one trapped cellular exon after (b).
24. The method of claim 22, further comprising identifying at least one trapped cellular exon after (c).
25. The method of claim 21, wherein the transgenic mouse comprising the vector is a somatic transgenic mouse.
26. The method of claim 21, wherein the transgenic mouse comprising the vector is a germ line transgenic mouse.
27. The method of claim 21, wherein the first exon sequence additionally encodes an internal ribosome entry site operatively positioned between said splice acceptor and an initiation codon of said first exon.
28. The method of claim 21, wherein the vector additionally comprises in the region between said polyadenylation sequence and said first promoter at least one of a transcription termination sequence, a 3 terminal exon, and a sequence encoding a self-cleaving RNA.
29. The method of claim 21, wherein the marker encoded by the first exon sequence of the vector is selected from a marker conferring antibiotic resistance, a marker conferring antibiotic sensitivity, an enzymatic marker, a recombinase, and a fluorescent marker.
30. The method of claim 29 wherein the marker confers neomycin resistance.
31. The method of claim 21, wherein the vector is selected from a viral vector and a retroviral vector.
32. The method of claim 23, wherein the identifying at least one trapped cellular exon comprises:
a) obtaining a chimeric transcript resulting from splicing of the second exon sequence to a third exon sequence, wherein the third exon sequence is from the genome of the ES cell;
b) reverse transcribing said chimeric transcript to produce a cDNA template; and
c) determining the polynucleotide sequence of the cDNA template.
33. The method of claim 24, wherein the identifying at least one trapped cellular exon comprises:
a) obtaining a chimeric transcript resulting from splicing of the second exon sequence to a third exon sequence, wherein the third exon sequence is from the genome of the transgenic mouse;
b) reverse transcribing said chimeric transcript to produce a cDNA template; and
c) determining the polynucleotide sequence of the cDNA template.
34. A method of making a transgenic mouse comprising a vector, comprising
a) introducing a vector into murine embryonic stem (ES) cells, wherein the vector comprises a 3 gene trap cassette, comprising in operable combination:
i) a promoter;
ii) an exon sequence located 3 from and expressed by said first promoter, said exon sequence not encoding an activity conferring antibiotic resistance; and
iii) a splice donor sequence located at the 3 end of said exon sequence;
wherein the vector does not encode a sequence that mediates the polyadenylation of an mRNA transcript encoded by said exon sequence;
b) selecting a murine ES cell that comprises the vector; and
c) making a transgenic mouse comprising the vector from the selected murine ES cell that comprises the vector.
35. The method of claim 34, wherein the vector from the selected murine ES cell that comprises the vector is non-homologously incorporated into the genome of at least one cell in the transgenic mouse.
36. The method of claim 35, further comprising identifying at least one trapped cellular exon after (b).
37. The method of claim 35, further comprising identifying at least one trapped cellular exon after (c).
38. The method of claim 34, wherein the transgenic mouse comprising the vector is a somatic transgenic mouse.
39. The method of claim 34, wherein the transgenic mouse comprising the vector is a germ line transgenic mouse.
40. The method of claim 34, wherein the exon sequence additionally encodes an internal ribosome entry site operatively positioned between said splice acceptor and an initiation codon of said exon sequence.
41. The method of claim 34, wherein the vector additionally comprises in the region between said polyadenylation sequence and said promoter at least one of a transcription termination sequence, a 3 terminal exon, and a sequence encoding a self-cleaving RNA.
42. The method of claim 34, wherein the exon sequence encodes a marker selected from an enzymatic marker, a recombinase, and a fluorescent marker.
43. The method of claim 42 wherein the marker is a fluorescent marker.
44. The method of claim 34, wherein the vector is selected from a viral vector and a retroviral vector.
45. The method of claim 36, wherein the identifying at least one trapped cellular exon comprises:
a) obtaining a chimeric transcript resulting from splicing of the exon sequence from the vector to a second exon sequence, wherein the second exon sequence is from the genome of the ES cell;
b) reverse transcribing said chimeric transcript to produce a cDNA template; and
c) determining the polynucleotide sequence of the cDNA template.
46. The method of claim 37, wherein the identifying at least one trapped cellular exon comprises:
a) obtaining a chimeric transcript resulting from splicing of the exon sequence from the vector to a second exon sequence, wherein the second exon sequence is from the genome of the transgenic mouse;
b) reverse transcribing said chimeric transcript to produce a cDNA template; and
c) determining the polynucleotide sequence of the cDNA template.