Table 1.

Features of Insertional Mutants

	No. of Transformants			Complementation with Strains
Mutants	Plasmid	+	−	305(nia1)	nit2	Insertion Localization	Distance to NIT2
AE1	pMN68	84	0	+	−	n.d.	n.d.
AE2	pMN68	69	0	+	−	n.d.	n.d.
AE9	pMN68	36	0	+	−	n.d.	n.d.
AE10	pMN68	104	0	+	−	n.d.	n.d.
AE20	pMN68	31	0	+	−	n.d.	n.d.
89.87	pNit2	42	0	+	−	9(324200↓)	0
177.1	pNit2	79	0	+	−	9(329046⊣)	2.5
	pMN68	78	0
229.13	pNit2	5	0	+	−	9(323980↓)	0
257.67	pNit2	6	0	+	−	9(304248⊢)	15
257.92

The five AE mutants were isolated after insertional mutagenesis using the pBle marker DNA (Lumbreras et al., 1998), and the other five mutants were isolated using the pAphVIII marker (Sizova et al., 2001). Transformation of mutants with the indicated plasmid DNA bearing the complete NIT2 gene was performed with 1 μg of DNA (+) or with no DNA (−) as a control (Kindle, 1990). Genetic complementation with the nit2 mutant was performed as detailed in Methods; −, no complementation. The insertion of the bleomycin resistance gene is unlinked to the Nit⁻ phenotype and was not determined (n.d.). Localizations of the paromomycin resistance gene insertions for the indicated mutants are indicated according to version 3 of the JGI at the Chlamydomonas genome server as follows: first number, number of scaffold; numbers in parentheses, position in the scaffold of the insertion; symbols ↓,⊢, and ⊣, insertion positions of the inserted plasmid DNA within the NIT2 gene, toward the right, or toward the left, respectively. Mutants 257.67 and 257.92 correspond to an identical insertion event. GenBank National Center for Biotechnology Information accession numbers for genomic sequences adjacent to the marker insertion are as follows: DQ385508, mutant 89.87; AY704202, mutant 177.1; DQ385509, mutant 229.13; and AY704210, mutant 257.67.