Alcohol dehydrogenase specific activities at different temperatures.

<p>*indicates that the difference is significant (P<0.05).</p><p>Mean ± SD from three independent experiments are shown.</p

Identification of the mutant with CotC-BmADH integration at <i>amyE</i> locus.

<p>(A) Analysis of amylase activity. CotC-BmADH mutant strains and <i>B. subtilis</i> 168 (trp-) wide type grew on the starch-containing LB plate before (1) and after (2) being stained by iodine. The integration of CotC-BmADH might disrupt <i>amyE</i> and made the strain amylase deficient, while the while wide type strain showed a big white halo around colony due the secretion of amylase. (B) Site-directed PCR analysis using different primer pairs. Marker, <i>λ</i> DNA digested by <i>Eco</i>T14I; W: <i>B. subtilis</i> 168 (trp-) wide typ; M: CotC-BmADH mutant; primer pairs used in PCR are labeled below agarose gel.</p

Cloning strategy.

<p>(A) The construction of integration plasmid pJS700-BmADH. The fragments <i>amyE</i> 5′ and <i>amyE</i> 3′ in plasmid are homologous to the upstream and downstream of the amylase gene in <i>B. subtilis</i> 168 (trp-), respectively; <i>Em</i>^r, erythromycin resistant site; <i>CotC</i>, a <i>B. subtilis</i> spore coat protein encoding gene. (B) The schematic integration of CotC-BmADH to <i>amyE</i> locus. Arrows indicate the positions of primer pairs used in the site-directed PCR for confirmation of the correct integration.</p

Alcohol dehydrogenase specific activities at different pHs.

<p>*indicates that the difference is significant (P<0.05).</p><p>Mean ± SD from three independent experiments are shown.</p

Relative remained activity of BmADH and CotC-BmADH after incubation at 37°C for 30 min.

<p>The activity was assayed at pH 9.0, 25°C. *indicates the difference is significant (P<0.05).</p

SDS-PAGE analysis of CotC-BmADH and Western blotting.

<p>(A) SDS-PAGE stained by coomassie-blue. (B) CotC-BmADH detected by BmADH specific antibody. Lane 1, <i>B. subtilis</i> 168 (trp-); lane 2, CotC-BmADH strain.</p

The Spatiotemporal expression profiles of BmTHYs.

<p>Western blot analysis of the expression levels of BmTHYs in different developmental stage (A) and in different tissues and organs (B). The mass of each lane’s total protein were 50 μg, and the sample were equalized. (A) Lane 1: egg; Lane 2: 1st instar; Lane 3: 2nd instar; Lane 4: 3rd instar; Lane 5: 4th instar; Lane 6: 5th instar; Lane 7: pupae; Lane 8: moth. (B) Lane 1: midgut; Lane 2: testis; Lane 3: ovary; Lane 4: head; Lane 5: fat body; Lane 6: hemolymph.</p

The expression pattern of BmTHYs in BmN cells infected with BmNPV.

<p>The BmN cells were infected with BmNPV and samples were collected at different hours to examine the expression of BmTHYs. The mass of each lane’s total protein were 30 μg, and the sample were equalized.</p