US 11,781,128 B2
Methods for producing hybrid polyketide synthase genes and polyketides
Richard Reumerman, Cambridge (GB); Matthew Alan Gregory, Cambridge (GB); Aleksandra Maria Wlodek, Cambridge (GB); and Divyang Jani, Cambridge (GB)
Assigned to ISOMERASE THERAPEUTICS LIMITED, Cambridge (GB)
Appl. No. 16/319,868
Filed by Isomerase Therapeutics Limited, Cambridge (GB)
PCT Filed Jul. 28, 2017, PCT No. PCT/GB2017/052221
§ 371(c)(1), (2) Date Jan. 23, 2019,
PCT Pub. No. WO2018/020272, PCT Pub. Date Feb. 1, 2018.
Claims priority of application No. 1613130 (GB), filed on Jul. 29, 2016; and application No. 1703657 (GB), filed on Mar. 7, 2017.
Prior Publication US 2020/0024595 A1, Jan. 23, 2020
Int. Cl. C12N 15/70 (2006.01); C12N 15/81 (2006.01); C12N 15/10 (2006.01)
CPC C12N 15/1055 (2013.01) [C12N 15/70 (2013.01); C12N 15/81 (2013.01)] 24 Claims
 
1. A method for producing a hybrid polyketide synthase (PKS) gene of a type 1 modular PKS in which one or more modules are deleted or inserted which comprises joining three or more sections of DNA encoding PKS genes or parts thereof no pair of which is contiguous in nature and creating two or more junctions between said sections of DNA (a) within DNA encoding a ketosynthase (KS) domain so that two KS domains KSn and KSn+1 are joined at a position in a region within those residues of the amino acid sequences of said KSn and KSn+1 that are aligned with amino acids 199-342 of SEQ ID NO 37 as well as amino acids 203-346 of SEQ ID NO 38; or (b) within DNA encoding an acyltransferase (AT) domain so that two AT domains ATn and ATn+1 are joined at a position in a region within those residues of the amino acid sequences of said ATn and ATn+1 that are aligned with amino acids 5-246 of SEQ ID NO 85 or amino acids 5-269 of SEQ ID NO 86 or (c) in DNA encoding the inter-domain region before the acyl carrier protein (ACP) domain so that two pre-ACP domain regions pre-ACPn and pre-ACPn+1 are joined at a position in a region within those residues of the amino acid sequences of said pre-ACPn and pre-ACPn+1 domains that are aligned with amino acids 184-268 of SEQ ID NO 39, amino acids 184-269 of SEQ ID NO 40 and amino acids 184-270 of SEQ ID NO 41.