. 2011 Jan 7;23(1):333–349. doi: 10.1105/tpc.110.078170

Table 2.

C. reinhardtii Strains Generated by Transformation

Recipient Strain	Transforming Plasmid	Transformed Strains
Nuclear transformation^a
mca1-6	plgMCA1-HA [1]^b	mH
mtF	plgMCA1-HA [1]^b	mHtF
tca1-8	pshTCA1-Fl [1]^c	CTCA1
Chloroplast transformation^d
mH	pΔpetA [2]	mH {ΔpetA}^e
mH	pWF_StopK [this study]	mH {petA_St}^e
mH	p5′dAf K [this study]	mH {5′dAf }^f
mH	pf₂₈₃St [3]	mH {f_Sol}^e ^g
mH	pf₃₁₂St [4]	mH {f₃₁₂St}^g
mH	pf₃₁₀St [4]	mH {f₃₁₀St}^g
mH	pf₃₀₇St [4]	mH {f₃₀₇St}^g
mH	pf₃₀₄M [4]	mH {f₃₀₄M}^h
mH	pf₃₀₇S [4]	mH {f₃₀₇S}^h
mH	pfΔK [4]	mH {fΔK}^h
mH	p5′dAf₃₀₇S [this study]	mH {5′dAf₃₀₇S}^f ^h
mH	pΔpetD [2]	mH {ΔpetD}^e
mHt	pΔpetA [2]	mHt {ΔpetA}^f
mHt	p5′dAfK [this study]	mHt {5′dAf}^f ⁱ
mHt	p5′dAf₃₀₇S [this study]	mHt {5′dAf₃₀₇S}^h ⁱ
tF	pΔpetA [2]	tF {ΔpetA}^e
mtF	pΔpetA [2]	mtF {ΔpetA}^f

References are as follows: 1 (Raynaud et al., 2007), 2 (Kuras and Wollman, 1994), 3 (Kuras et al., 1995a); 4 (Choquet et al., 2003), and 5 (Loiselay et al., 2008).

All recipient strains were nonphotosynthetic, and transformants were selected for photoautotrophy on minimum medium (Harris, 1989) under high light (200 μE·m⁻²·s⁻¹).

Plasmid DNA was linearized with XbaI that cuts upstream of the MCA1 initiation codon before transformation.

Plasmid was cut with XbaI and SfiI, and the 5461-bp fragment that encodes the C-terminal domain of TCA1 was used for transformation after gel purification.

All recipient strains were spectinomycin sensitive. Transformed strains were selected for resistance to spectinomycin (100 μg·mL⁻¹) under low light (5 μE·m⁻²·s⁻¹) and subcloned in darkness until they reached homoplasmy, unless otherwise indicated.

Homoplasmy was deduced from the loss of photoautotrophic growth capacity.

Homoplasmy was assessed by RNA gel blot experiments.

Homoplasmy was assessed by protein immunoblot analysis for the expression of truncated versions of cytochrome f.

The presence of the f₃₀₇S substitution was screened by restriction fragment length polymorphism analysis of PCR products amplified with primers Test_petA_For and Test_petA_Rev, as described by Choquet et al. (2003 [see Supplemental Table A]).

ⁱ

Strains were subcloned under dim light (25 μE·m² ·s⁻¹) as recipient strains were nonphotosynthetic mutants, whereas the transformed strains regained photoautrophic growth capability.