	US 11,807,901 B2
	Methods and kits for using recombinant microorganisms as direct reagents in biological applications
Andrew Ellington, Austin, TX (US); Sanchita Bhadra, Austin, TX (US); Jared Ellefson, Tucson, AZ (US); Jimmy Gollihar, Boston, MA (US); Arti Pothukuchy, Austin, TX (US); Michelle Byrom, Austin, TX (US); and Raghav Shroff, Austin, TX (US)
Assigned to BOARD OF REGENTS, THE UNIVERSITY OF TEXAS SYSTEM, Austin, TX (US)
Appl. No. 16/764,642
Filed by BOARD OF REGENTS, THE UNIVERSITY OF TEXAS SYSTEM, Austin, TX (US)
PCT Filed Nov. 15, 2018, PCT No. PCT/US2018/061247 § 371(c)(1), (2) Date May 15, 2020, PCT Pub. No. WO2019/099644, PCT Pub. Date May 23, 2019.
Claims priority of provisional application 62/586,606, filed on Nov. 15, 2017.
Prior Publication US 2020/0399679 A1, Dec. 24, 2020
Int. Cl. C12Q 1/686 (2018.01)

CPC C12Q 1/686 (2013.01)

12 Claims

1. A method of utilizing an enzyme in a nucleic acid manipulation process, the method comprising:

a. transforming a microorganism with a non-native enzyme;

b. inducing expression of the enzyme in the microorganism, thereby producing the non-native enzyme;

c. dehydrating or freeze-drying the microorganism without using an excipient;

d. rehydrating the microorganism of step c);

e. adding the microorganism of step d) directly to a non-naturally occurring nucleic acid manipulation process, thereby obtaining a non-purified cellular reagent, including the non-native enzyme, wherein the non-native enzyme is not purified from the microorganism prior to addition to the nucleic acid manipulation process;

f. carrying out the nucleic acid manipulation process using the enzyme.