INCORPORATION OF UNNATURAL NUCLEOTIDES AND METHODS THEREOF
Disclosed herein are methods, compositions and kits for the synthesis of proteins which comprises unnatural amino acids that utilize a mutant tRNA.
1 .- 23 . (canceled)
24 . An E. coli cell comprising:
(a) a nucleoside triphosphate transporter from Phaeodactylum tricornutum;
(b) an mRNA comprising an unnatural base comprising a structure selected from
(c) a tRNA comprising an unnatural base comprising a structure selected from
and
(d) an aminoacyl tRNA synthetase derived from an organism selected from the group consisting of Methanosarcina barkeri and Methanococcus jannaschii.
25 . The E. coli cell of claim 24 , wherein the cell further comprises an oligonucleotide encoding the mRNA.
26 . The E. coli cell of claim 24 , wherein the cell further comprises an oligonucleotide encoding the tRNA.
27 . The E. coli cell of claim 24 , wherein the cell further comprises an oligonucleotide encoding the aminoacyl tRNA synthetase.
28 . The E. coli cell of claim 24 , wherein the cell further comprises a single oligonucleotide encoding the mRNA and the tRNA.
29 . The E. coli cell of claim 24 , wherein the cell further comprises a single oligonucleotide encoding the mRNA, the tRNA, and the aminoacyl tRNA synthetase.
30 . The E. coli cell of claim 24 , wherein the mRNA comprises one or more codons selected from AXC and GXC, wherein X is the unnatural base.
31 . The E. coli cell of claim 30 , wherein the mRNA comprises the codon AXC.
32 . The E. coli cell of claim 31 , wherein the unnatural base comprises the structure
33 . The E. coli cell of claim 31 , wherein the unnatural base comprises the structure
34 . The E. coli cell of claim 30 , wherein the mRNA comprises the codon GXC.
35 . The E. coli cell of claim 34 , wherein the unnatural base comprises the structure
36 . The E. coli cell of claim 34 , wherein the unnatural base comprises the structure
37 . The E. coli cell of claim 24 , wherein the aminoacyl tRNA synthetase is derived from Methanosarcina barkeri.
38 . The E. coli cell of claim 36 , wherein the mRNA comprises an unnatural base comprising the structure
and the tRNA comprises an unnatural base comprising the structure
39 . The E. coli cell of claim 37 , wherein the mRNA comprises one or more codons selected from AXC and GXC, wherein X is the unnatural base.
40 . The E. coli cell of claim 24 , wherein the aminoacyl tRNA synthetase is derived from Methanococcus jannaschii.
41 . The E. coli cell of claim 39 , wherein the mRNA comprises an unnatural base comprising the structure
and the tRNA comprises an unnatural base comprising the structure
42 . The E. coli cell of claim 40 , wherein the mRNA comprises one or more codons selected from AXC and GXC, wherein X is the unnatural base.
43 . The E. coli cell of claim 24 , wherein the tRNA is derived from an organism selected from the group consisting of Methanosarcina barkeri and Methanococcus jannaschii.