Figure 6.

Detection of the RBR1 Gene and RBR1 Splice Variants in Transgenic Males Transformed with the Female RBR1 Gene.

(A) DNA gel blot hybridization (standard conditions) of genomic DNA from parent male 281-1 (control) and transgenic males transformed with the female RBR1 gene. Transformants T175-3-2, T175-4-6, and T175-6-1 are descendants of 281-1 that have been transformed with both the nitA gene and the female RBR1 gene. The DNA was digested with AccI. The blot was probed using a 754-bp RBR1 fragment.

(B) Relationship between RBR1 splice variants 1 to 4 (var1 to var4) in wild-type females and in transgenic males expressing the female RBR1 gene. Error bars refer to the sd of the y value. All real-time RT-PCR experiments were performed in triplicate from technical repeats. For calculation of all relative expression levels, the expression level of splice variant 1 in wild-type females was used as the reference point (=100). For isolation of mRNA from each strain, asynchronous, vegetatively grown cultures containing a mixture of the different developmental stages were used.

(B1) Distribution of splice variants in the female wild-type strain Eve10.

(B2) to (B4) Expression of the Volvox RBR1 gene and distribution of splice variants in transgenic males transformed with the female RBR1 gene.

(B2) Male transformant T175-3-2.

(B3) Male transformant T175-4-6.

(B4) Male transformant T175-6-1.

(C) Phenotypes of male TRef1 (control), a transgenic descendant of 281-1 that has been transformed only with the nitA selectable marker gene, and of transgenic males expressing the female RBR1 gene.

(C1) Male TRef1 (control).

(C2) Male transformant T175-3-2.

(C3) Male transformant T175-4-6.

(C4) Male transformant T175-6-1.