1. A nucleic acid construct comprising a regulatory element and a FatB gene expressing an active acyl-ACP thioesterase operable to produce an altered fatty acid profile in an oil produced by a cell expressing the nucleic acid construct, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 5 of Table 1a, the sequence having at least 94.6% sequence identity with each of SEQ ID NOs: 88, 82, 85, and 103, and optionally wherein the fatty acid of the oil is enriched in C8 and C10 fatty acids.
2. A nucleic acid construct comprising a regulatory element and a FatB gene expressing an active acyl-ACP thioesterase operable to produce an altered fatty acid profile in an oil produced by a cell expressing the nucleic acid construct, wherein the FatB gene expresses a protein having an amino acid sequence falling within one of clades 1-12 of Table 1a.
3. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 1 of Table 1a, the sequence having at least 85.9% sequence identity with each of SEQ ID NOs: 19, 161, 22, and 160, and optionally wherein the fatty acid of the oil is enriched in C14 and C16 fatty acids.
4. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 2 of Table 1a, the sequence having at least 89.5% sequence identity with each of SEQ ID NOs: 134-136, 132, 133, 137, 124, 122, 123, 125, and optionally wherein the fatty acid of the oil is enriched in C12 and C14 fatty acids.
5. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 3 of Table 1a, the sequence having at least 92.5% sequence identity with each of SEQ ID NOs: 126 and 127, and optionally wherein the fatty acid of the oil is enriched in C12 and C14 fatty acids.
6. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 4 of Table 1a, the sequence having at least 83.8% sequence identity with SEQ ID NO: 79, and optionally wherein the fatty acid of the oil is enriched in C12 and C14 fatty acids.
7. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 6 of Table 1a, the sequence having at least 99.9% sequence identity with each of SEQ ID NOs: 111 and 110, and optionally wherein the fatty acid of the oil is enriched in C10 fatty acids.
8. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 7 of Table 1a, the sequence having at least 89.5% sequence identity with each of SEQ ID NOs: 73, 106, 185, 172, 171, 173, 174, and optionally wherein the fatty acid of the oil is enriched in C10 and C12 fatty acids.
9. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 8 of Table 1a, the sequence having at least 85.9% sequence identity with each of SEQ ID NOs: 112, 113, 142, 145, 143, 144, 139, 140, 138, 141, and optionally wherein the fatty acid of the oil is enriched in C12 and C14 fatty acids.
10. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 9 of Table 1a, the sequence having at least 83.8% sequence identity with each of SEQ ID NOs: 187-189, and optionally wherein the fatty acid of the oil is enriched in C12 and C14 fatty acids.
11. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 10 of Table 1a, the sequence having at least 95.9% sequence identity with each of SEQ ID NOs: 147, 149, 146, 150, 152, 151, 148, 154, 156, 155, 157, 108, 75, 190, 191, and 192, and optionally wherein the fatty acid of the oil is enriched in C14 and C16 fatty acids.
12. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 11 of Table 1a, the sequence having at least 88.7% sequence identity with SEQ ID NO: 121, and optionally wherein the fatty acid of the oil is enriched in C14 and C16 fatty acids.
13. The nucleic acid construct of claim 2, wherein the FatB gene expresses a protein having an amino acid sequence falling within Glade 12 of Table 1a, the sequence having at least 72.8% sequence identity with each of SEQ ID NOs: 129 and 186, and optionally wherein the fatty acid of the oil is enriched in C16 fatty acids.
14. An isolated nucleic acid or recombinant DNA construct comprising a nucleic acid, wherein the nucleic acid has at least 70% sequence identity to any of SEQ ID NOS: 2, 3, 5, 6, 8, 9, 11, 12, 14, 15, 17, 18, 20, 21, 23, 24, 26, 27, 29, 30, 32, 33, 35, 36, 38, 39, 41, 42, 44, 45, 47, 48, 50, 51, 53, 54, 56, 57, 59, 60, 62, 63, 65, 66, 68, 69, 71, 72, 74, 76, 78, 80, 81, 83, 84, 86, 87, 89, 90, 92, 93, 95, 96, 98, 99, 101, 102, 104, 105, 107, 109 or any equivalent sequences by virtue of the degeneracy of the genetic code.
15. An isolated nucleic acid sequence encoding a protein or a host cell expressing a protein having at least 70% sequence identity to any of SEQ ID NOS: 1, 4, 7, 10, 13, 16, 19, 22, 25, 28, 31, 34, 37, 40, 43, 46, 49, 52, 55, 58, 61, 64, 67, 70, 73, 75, 77, 79, 82, 85, 88, 91, 94, 97, 100, 103, 106, 108, 110-192 or a fragment thereof having acyl-ACP thioesterase activity.
16. The isolated nucleic acid of claim 15, wherein, the protein has acyl-ACP thioesterase activity operable to alter the fatty acid profile of an oil produced by a recombinant cell comprising that sequence.
17. A method of producing a recombinant cell that produces an altered fatty acid profile, the method comprising transforming the cell with a nucleic acid according to claim 2.
18. A host cell produced by the method of claim 17.
19. The host cell of claim 18, wherein the host cell is selected from a plant cell, a microbial cell, and a microalgal cell.
20. A method for producing an oil or oil-derived product, the method comprising cultivating a host cell of claim 18, and extracting oil produced thereby, optionally wherein the cultivation is heterotrophic growth on sugar.
21. The method of claim 20, further comprising producing a fatty acid, fuel, chemical, or other oil-derived product from the oil.
22. An oil produced by the method of claim 20, optionally having a fatty acid profile comprising at least 20% C8, C10, C12, C14 or C16 fatty acids.
23. An oil-derived product produced by the method of claim 21.
24. The oil of claim 23, wherein the oil is produced by a microalgae and optionally, lacks C24-alpha sterols.
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. A hydraulic rescue tool comprising:
a) a hydraulic cylinder having a housing, a piston rod, and hydraulic controls, with said piston rod movably disposed within said housing, thus providing linear driving means for the rescue tool,
b) a cylinder yoke rotatably attached to the hydraulic cylinder housing, said cylinder yoke having means for manipulating the tool by hand, with rotation of the cylinder yoke relative to the cylinder, thereby preventing injury to an operator in the event that the tool binds and torques, said cylinder yoke further having a pair of pivotal links attached thereunto,
c) an implement unit detachably disposed onto said piston rod and said pivotal links for cutting or spreading metal, and
d) a plurality of quick release pins attaching said implement unit in concert with said piston rod and said pivotal links, thus providing a rapid release of the implement unit from the piston rod and links for replacement or repair.
2. The hydraulic rescue tool as recited in claim 1 wherein said hydraulic cylinder further having internal grooves and said cylinder yoke having mating external lands, and wherein said piston rod having a wet seal between the cylinder and the piston rod that permits both the yoke and the piston rod to revolve in unison from the hydraulic cylinder.
3. The hydraulic rescue tool as recited in claim 2 further comprising a drag control and locking mechanism that is defined as a spring-loaded ball detent disposed within said cylinder yoke, with the ball of the detent engaging the hydraulic cylinder adjacent to said internal grooves, such that tightening the mechanism increases drag to a point of locking engagement, thereby disabling rotation of the cylinder relative to the yoke.
4. The hydraulic rescue tool as recited in claim 1 wherein said implement unit further comprises a cutter for severing material such as found in vehicle crashes when a vehicle is deformed and the material must be quickly cut away to rescue trapped persons.
5. The hydraulic rescue tool as recited in claim 4 wherein said cutter further comprises a two-piece cutter clevis attached to said piston rod on one end, and a pair of cutting blades attached to the cutter clevis on the other, such that when the piston rod is retracted within the hydraulic cylinder housing the blades rotate towards each other on a common pivot in scissor fashion, thus forming a cutting surface therebetween.
6. The hydraulic rescue tool as recited in claim 5 wherein each cutting blade further comprises a primary blade and a backup plate, with the primary blade having a sharp cutting edge and the backup plate having significant structural integrity for reinforcing the primary blade and to preclude the blade breaking and flying into pieces.
7. The hydraulic rescue tool as recited in claim 6 wherein said backup plate is fabricated by a process selected from the group consisting of: machining, casting, forging a single element, and a two-piece construction with the pieces welded together.
8. The hydraulic rescue tool as recited in claim 6 wherein said primary blade and said backup plate are attached together by a process selected from the group consisting of: a plurality of rivets, a plurality of bolts, and brazing together in a furnace.
9. The hydraulic rescue tool as recited in claim 5 wherein said cutter further comprises a plurality of thrust washers and a plurality of roller bearings that are spaced between said a pair of cutting blades and held jointly with a pivot bolt and a nut, thereby forcing the cutting blades tightly together.
10. The hydraulic rescue tool as recited in claim 1 wherein said implement unit further comprises a spreader that is defined as a pair of spreader arms and a spreader clevis which attaches both arms in such a manner that they pivotally arc into a spread-open position and intimately engage with each other in a closed position.
11. The hydraulic rescue tool as recited in claim 10 wherein said spreader clevis further comprises a spreader pivot bolt connecting each spreader to said spreader clevis.
12. The hydraulic rescue tool as recited in claim 1 wherein said pivotal links further having a cavity therein along a longitudinal surface that provides a bendable section for failing in a safe manner in the event of over pressurization of the hydraulic cylinder.
13. The hydraulic rescue tool as recited in claim 12 wherein said pivotal links are constructed of an aluminum material.
14. The hydraulic rescue tool as recited in claim 1 wherein said piston rod further having an extending end and an internal end, with said internal end having a diameter rebated at least two times smaller than the extended end.
15. A rescue tool comprising:
a) linear driving means having a pair of pivotal links and a piston rod, with said piston rod movably providing a pushing and a pulling action for the rescue tool,
b) an implement unit detachably connected to said piston rod and said pivotal links for cutting and spreading metal during a rescue,
c) a plurality of quick release pins attaching said implement unit in concert with said piston rod and said pivotal links, thereby providing a rapid release of the implement unit from the piston rod and pivotal links for quick replacement or repair, and
d) said pivotal links having a cavity therein along a longitudinal surface that provides a bendable section for failing in a safe manner in the event of excessive force created by said linear driving means.
16. A hydraulic rescue tool comprising:
a) a hydraulic cylinder, having a cylinder housing with a piston rod and a cylinder yoke rotatably attached to said cylinder housing, thus providing a pushing and a pulling action for the rescue tool,
b) said hydraulic cylinder housing having internal grooves, said cylinder yoke having mating external lands, and said piston rod having a wet seal between the cylinder housing and the piston rod, and
c) said cylinder yoke having means for manipulating the tool by hand, which permits both the cylinder yoke and piston rod to revolve in unison relative to the hydraulic cylinder housing, thereby preventing injury to an operator in the event that the tool binds and torques.
17. A rescue tool comprising:
a) linear driving means having a piston rod and pivotal links providing a pushing and a pulling action for the rescue tool,
b) an implement unit connected to said linear driving means for cutting and spreading metal during a rescue,
c) said implement unit comprises a cutter for severing material such as found in vehicle crashes when the vehicle is deformed and the material must be quickly cut away to rescue trapped persons,
d) said cutter further comprises a pair of cutting blades such that when the linear driving means is retracted the cutting blades rotate towards each other on a common pivot in scissor fashion, thus forming a cutting surface therebetween,
e) each cutting blade comprises a primary blade and a backup plate, with the primary blade having a sharp cutting edge and the backup plate having significant structural integrity,
f) said backup plate fabricated by a process selected from the group consisting of: machining, casting, forging a single element, and two-piece construction with the pieces welded together,
g) a plurality of quick release pins attaching said implement unit in concert with said piston rod and said pivotal links, thus providing a rapid release of the implement unit from the piston rod and links for replacement or repair, and
h) said pivotal links having a cavity therein along a longitudinal surface that provides a bendable section for failing in a safe manner in the event of excessive force created by said linear driving means.
18. A rescue tool comprising:
a) linear driving means having a pair of pivotal links, said linear driving means providing a pushing and a pulling action for the rescue tool, and
b) said pivotal links having a cavity therein along a longitudinal surface that provides a bendable section for failing in a safe manner in the event of excessive force created by said linear driving means.
19. A hydraulic rescue tool comprising:
a) a hydraulic cylinder, which has a hydraulic housing and a piston rod, that provides a pushing and a pulling action for the rescue tool, and
b) said piston rod having an extending end and an internal end, with said internal end having a diameter rebated at least two times smaller than the piston extending end.