Fig. 2

Agar gel analysis. a Verification of tri3 deletion. Numbers from 1 to 10 and 12 to 21 refer to the arbitrarily chosen transformants. Numbers 11 and 22 refer to the wild strain. The numbers and their corresponding transformants are as follows: 1 and 12, Δtri3-1; 2 and 13, Δtri3-2; 3 and 14, Δtri3-3; 4 and 15, Δtri3-4; 5 and 16, Δtri3-5; 6 and 17, Δtri3-6; 7 and 18, Δtri3-7; 8 and 19, Δtri3-8; 9 and 20, Δtri3-9; and 10 and 21, Δtri3-10. Lanes 2, 5, 6, and 10 did not show the expected band of 1.1 kb. This result indicated that tri 3 was deleted in Δtri3-2, Δtri3-5, Δtri3-6, and Δtri3-10, respectively. Furthermore, Δtri3-2, Δtri3-5, Δtri3-6, and Δtri3-10 (lanes 13, 16, 17, and 21, respectively) showed the expected band of 3.6 kb (reduced by approximately 0.7 kb). The ORF of the tri3 was replaced successfully by the hygromycin cassette in these mutants. b Verification of tri11 deletion. Numbers from 1 to 9 and 11 to 19 refer to the arbitrarily chosen transformants, while numbers 10 and 20 refer to the wild strain. The numbers and their corresponding transformants are as follows: 1 and 11, Δtri11-1; 2 and 12, Δtri11-2; 3 and 13, Δtri11-3; 4 and 14, Δtri11-4; 5 and 15, Δtri11-5; 6 and 16, Δtri11-6; 7 and 17, Δtri11-7; 8 and 18, Δtri11-8; and 9 and 19, Δtri11-9. Lanes 1, 4, 5, and 7 did not show the expected band of 1.1 kb. This result indicated that tri11 was deleted in Δtri11-1, Δtri11-4, Δtri11-5, and Δtri11-7, respectively. Furthermore, Δtri11-1, Δtri11-4, Δtri11-5, and Δtri11-7 (lanes 11, 14, 15, and 17, respectively) showed the expected band of 3.6 kb (reduced by approximately 0.7 kb). The ORF of the tri11 was replaced successfully by the hygromycin cassette in these mutants. c Verification of tri4 deletion. Numbers from 1 to 9 and 11 to 19 refer to the arbitrarily chosen transformants, while numbers 10 and 20 refer to the wild strain. The numbers and their corresponding mutants are as follows: 1 and 11, Δtri4-1; 2 and 12, Δtri4-2; 3 and 13, Δtri4-3; 4 and 14, Δtri4-4; 5 and 15, Δtri4-5; 6 and 16, Δtri4-6; 7 and 17, Δtri4-7; 8 and 18, Δtri4-8; and 9 and 19, Δtri4-9. Lanes 3, 4, 6, 8, and 9 did not show the expected band of 1.1 kb. This result indicated that tri4 was deleted in Δtri4-3, Δtri4-4, Δtri4-6, Δtri4-8, and Δtri4-9, respectively. Furthermore, Δtri4-3, Δtri4-4, Δtri4-6, Δtri4-8, and Δtri4-9 (lanes 13, 14, 16, 18, and 19, respectively) showed the expected band of 3.6 kb (reduced by approximately 0.7 kb). The ORF of the tri4 was replaced successfully by the hygromycin cassette in these mutants