Selection markers for transformation of the sequenced reference monokaryon Okayama 7/#130 and homokaryon AmutBmut of Coprinopsis cinerea

Table 3 Transformations of C. cinerea FA2222 (trp1.1,1.6) with plasmid pBD5 alone or, using same batches of protoplasts, in combination with other non-directly selectable vectors

Plasmid(s)	trp1⁺-selected transformants collected on					Total transformants	Ratio of clones
Plasmid(s)	1st day	2nd day	3rd day	4th day	5th day	Total transformants	Ratio of clones
Experiment 1
pBD5	13	8	12	3	2	38	1.0
pBD5 + pYSK7*	30 (8)	20 (13)	32 (7)	9 (2)	4 (2)	95 (32)	2.5
pBD5 + pDB3	32	13	25	6	3	79	2.1
pBD5 + pPAB1-2	18	17	17	7	2	61	1.6
pBD5 + pCcAde8	34	27	11	4	2	78	2.1
Experiment 2
pBD5	46	38	28	12	11	135	1.0
pBD5 + pYSK7*	94 (31)	89 (22)	68 (27)	15 (12)	5 (3)	271 (95)	2.0
pBD5 + pDB3	69	52	53	15	12	201	1.5
pBD5 + pPAB1-2	76	78	49	28	14	245	1.8
pBD5 + pCcAde8	100	114	90	26	14	344	2.5

*Date in brackets indicate clones expressing laccases as deduced from stained halos around their colonies. Non-producers of laccase did not stain the agar. Random subsets of unstained pBD5 and of staining pBD5 + pYSK7 clones from both experiments were further tested in liquid fermentations

ISSN: 2054-3085