Abstract: The present disclosure provides methods for incorporating at least one non-natural amino acid into a polypeptide using a cell-free protein synthesis which includes a cell-free extract and is deficient in endogenous tRNA. The methods include providing the synthesis system with at least one non-natural amino acid, at least one orthogonal tRNA and one orthogonal aminoacyl-tRNA synthetase which aminoacylates the corresponding orthogonal tRNA with the non-natural amino acid. The methods may also include removing the endogenous tRNA in the cell-free protein synthesis system by treating the synthesis system with a ribonuclease to degrade the endogenous tRNA and providing the synthesis system with a minimal set of tRNAs for one or more natural amino acids and the corresponding amino-acyl tRNA synthetases, such that the lack of some tRNA will enable unique codons to encode for non-natural amino acid incorporation with no or a minimal amount of completion from the tRNA present.

Abstract: A novel membrane preparation process via chemical vapor deposition (CVD) deposits a film on a microporous ceramic support at a low temperature (300.degree. to 550.degree. C.) with organic metallic compounds as precursors and then converts the film into porous oxide at a higher temperature (350.degree. C. to 800.degree. C., preferably 450.degree. C. to 600.degree. C.) to increase its permeability and stability at a given process application temperature. The novel membrane is used for gas separations at an elevated temperature (300.degree. C. to 800.degree. C.). The novel membrane can be used as a catalytic membrane reactor which selectively removes one of the reaction products, thereby enhancing the conversion of the reaction. The ultrafiltration membrane provides gaseous mixtures separation media having high permeance and superior hydrogen selectivity.