Valuation of agro-industrial wastes as substrates for heterologous production of α-galactosidase

Table 4 Estimated copy number of MEL1His-2 integrative cassette by relative quantification

Strain	Ct_{MEL1; n}	Ct_{TAF10; n}	∆Ct ^a_n (Ct_{MEL1; n} − Ct_{TAF10; n})	∆∆Ct (∆Ct_n − ∆Ct_c)	Copy number^b
Strain	Ct_{MEL1; n}	Ct_{TAF10; n}	∆Ct ^a_n (Ct_{MEL1; n} − Ct_{TAF10; n})	∆∆Ct (∆Ct_n − ∆Ct_c)	2^−∆∆Ct	(1 − E)^−∆∆Ct
KGM2^c	22.24 ± 0.43	24.24 ± 0.52	− 1.99 ± 0.41	− 0.01	1.01	1.01
KGM8	16.56 ± 0.29	19.51 ± 0.31	− 3.19 ± 0.24	− 1.20	2.31	2.45
KGM10	19.37 ± 0.32	22.91 ± 0.22	− 3.53 ± 0.34	− 1.55	2.94	3.17
KGM11	16.58 ± 0.45	19.27 ± 0.39	− 2.68 ± 0.43	− 0.70	1.62	1.68
KGM12	17.26 ± 0.12	21.14 ± 0.23	− 3.88 ± 0.23	− 1.89	3.71	3.97
KGM19	19.61 ± 0.20	22.52 ± 0.54	− 2.91 ± 0.21	− 0.92	1.90	1.99
KGM21	17.55 ± 0.62	21.42 ± 0.22	− 3.86 ± 0.47	− 1.88	3.68	4.08
KGM25	19.01 ± 0.16	22.05 ± 0.41	− 3.03 ± 0.44	− 1.05	2.07	2.18
KGM33	18.33 ± 0.32	21.65 ± 0.66	− 3.31 ± 0.12	− 1.28	2.43	2.59

^aCalculated from three extractions of genomic DNA samples, n = 3 ± SD
^b∆Ct_c (calibrator gene ∆Ct) and E (PCR efficiency) were obtained experimentally of the standard curves calculated from the serial tenfold dilutions of the genomic DNA (10 ng/µL) of the calibrator strain (∆Ct_c = 1.98 ± 0.38; E = 1.12; n = 4 ± SD; average values were evaluated by t-test, p-value < 0.05)
^cKGM2 was the single copy strain used as control of the calibrator to validate that it is not affected by experimental treatment

ISSN: 1475-2859