Patent SG-11201804966V-A (SG)
Title: Gene cassette for homologous recombination knock-out in yeast cells
Inventor: CHEN XIANZHONG
IPC: C12N15/81
Filed: 2015-12-17
Abstract: b) .6 kb-URA3 PCR product Tramsformation 'MCA 0_3 kb-LA Oa 0.3 kb-RA... Integration 1.6 kb - URA3 03 kb -LA Oa .CAT2n(7,1 0.3 kb-RA chromosome- 'CAT2udR C ATR 0:3 kb-LA gda 0.3 kb-RA :'• ' ' 4 , Excision (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2017/101060 Al 22 June 2017 (22.06.2017) WIPO I PCT (51) International Patent Classification: C12N 15/81 (2006.01) CJ2R 1/74 (2006.01) C12N 15/60 (2006.01) (21) International Application Number: PCT/CN2015/097675 (22) International Filing Date: 17 December 2015 (17.12.2015) (25) Filing Language: English (26) Publication Language: English (71) Applicant: EVONIK DEGUSSA (CHINA) CO., LTD. [CN/CN]; 55 Chundong Road, Xinzhuang Industry Park, Shanghai 201108 (CN). (72) Inventors: CHEN, Xianzhong; Lihu Jiayuan 46-601, Lihu Road 1800, Wuxi, Jiangsu 214122 (CN). ZHANG, Lihua; Zhangdazhuang, Ganglidian Township, Xixian County, Xinyang, Henan 464314 (CN). XIANG, Zheng; Yiquany- uan 25-501, Rongxiang Street, Binhu District, Wuxi, Ji- angsu 214000 (CN). SHEN, Wei; Wankemeili City 71- 403, Zhouxin Road, Wuxi, Jiangsu 214122 (CN). LI, Li; 1517 Huqingping Road, House 129, Shanghai 201702 (CN). POETTER, Markus; 658 Gaojing Road, House #85, Shanghai 201702 (CN). (74) Agent: CHINA PATENT AGENT (H.K.) LTD.; 22/F., Great Eagle Center, 23 Harbour Road, Wanchai, Hong Kong (CN). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Published: with international search report (Art. 21(3)) [Continued on next page] (54) Title: GENE CASSETTE FOR HOMOLOGOUS RECOMBINATION KNOCK-OUT IN YEAST CELLS 1111111111111101110101011111010111110 III 0101111101011011001111111111111110111111 (57) : A gene cassette for the knock-out of at least on target gene in yeast cells, the cassette comprises: (a) a URA3 gene as a marker gene; (b) at least one gene disruption auxiliary (gda) se- quence; and (c) the upstream and down- stream sequences of the target gene, wherein the gda sequence is a sequence having at least 300 to 600 consective residues of the nucleotide sequence of SEQ ID NO: 39. 0.3 kb-LA gda 0,3 kb - RA giSiV i • \ \ \ cAmdu ?ndk 0.3 kb-LA gda 0.3 kb- W O 20 17 / 10 1060 Al FIGURE 2 WO 2017/101060 Al 11111111111111011101111111111111111101111111111111111111111111111110N1011110111111 — with sequence listing part of description (Rule 5.2(a))