3 results about "Transport activity" patented technology

The invention discloses a transporter mutant capable of efficiently transporting non-natural nucleoside triphosphate. The molecular structure of a truncated body PtNTT2 (66-575) of a nucleoside triphosphate transporter PtNTT2 is analyzed, the PtNTT2 (66-575) is taken as an evolution starting point, key amino acid residues: 271th and 474th amino acids are selected, site-specific saturated mutation libraries are respectively constructed, and screening is carried out aiming at the transport activities of different non-natural nucleoside triphosphate, so that the site-specific saturated mutation libraries can be used for screening the transport activities of different non-natural nucleoside triphosphate. The mutant capable of efficiently transferring non-natural nucleoside triphosphate (dCNTMOTP, dNaMTP and NaMTP) is obtained by adopting the method provided by the invention. The mutant disclosed by the invention can be used for high-efficiency cell transmembrane delivery of various non-natural nucleoside triphosphates, the substrate range of nucleoside triphosphate transporter is expanded, and a foundation is laid for construction of semi-synthetic life bodies for efficiently expanding genetic letters.

The present application belongs to the technical field of sugar transporter activity detection, and particularly relates to a method for detecting the transport activity of vacuole membrane sugar transporter. The method comprises the following steps: constructing a plant expression vector for expressing the vacuole membrane sugar transporter, transforming the expression vector into the fruit of a melon at the swelling stage to obtain transgenic melon fruit; culturing the transgenic melon fruit until the vacuole membrane sugar transporter is expressed, and then obtaining protoplasts by enzymolysis; adding the protoplasts into a sugar fluorescent analogue culture solution, incubating for a period of time, and then detecting the fluorescence signal of the vacuoles in the protoplasts, and evaluating the transport activity of the vacuole membrane sugar transporter according to the fluorescence intensity. The method of the present application is simple to operate, high in efficiency, independent of isotopes, and capable of detecting the transport activity of both endogenous and exogenous vacuole sugar transporters, while maintaining the characteristics of the plant vacuole itself, reducing the interference of the background signal in the vacuole, and improving the accuracy and sensitivity of the analysis of the transport activity of the plant vacuole membrane sugar transporter.

This method for measuring transporter activity comprises: exposing a membrane vesicle having a vacuolar ATPase and a vesicular nucleotide transporter to an anion which is labeled with a radionuclide and is different from adenosine triphosphate, in a reaction solution containing adenosine triphosphate and magnesium ions; and measuring the transport activity of the anion into the membrane vesicle on the basis of the radiation of the radionuclide.