Revision as of 00:29, 20 October 2016

Team FAFU-CHINA

Proof

1.Introduction

In our project, we transform theexpression vector of Cry and Cyt toxins. Based on the toxin groups,we construct six various toxins express devices as Figure1. To compare the toxicities, we also tested the toxicity of Bacillus thuringiensis strain LLP229.

2.Material, Condition and Method

The larvae of Ae. albopictus are feeding in the Key Laboratory of Biopesticideand Chemical Biology. The generation of Ae.albopictus is 11. The LLP229strain of Bacillus thuringiensis isprovided by Key Laboratory of Biopesticide and Chemical Biology. The concentration of LLP229is recorded in eachtable. The CC530 and FA479 strains of Chlamydomonas reinhardtii and TAP mediumare bought from the Aquatic Species Research Institute in Wuhan, Hubei. The concentration of Chlamydomonasreinhardtii is recorded ineach table. We contrasted various toxins groupexpression vectors and transform them into Chlamydomonasreinhardtii as following groups: Cry4Aa+2A+Cyt1,Cry4Aa+2A+Cyt2, Cry10Aa+2A+Cyt1, Cry10Aa+2A+Cyt2, Cry11Aa+2A+Cyt1,Cry11Aa+2A+Cyt2. The toxicity analysis condition is in anenvironmentally controlled room (T=28±1 °C, RH=85%±5%, L: D=14:10). Bioassay of variousconcentrations were assayed against 30 third-instar larvae in 20 ml of dechlorinatedwater. Each group was replicated 3 times. The level of mortality was recordedafter 48 hours. Then calculated the corrected mortality. The bioassay was thenrepeated such that a concentration range yielding 10–90% mortality was obtainedand the 50% lethal concentration (LC₅₀) was calculated. Table 1.1 Toxicity of LLP29 to A. albopictus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	30	30	100
2	30	30	100
3	30	30	100
Control	30	0	0

Note. Thenconcentration of LLP29 is 2.16 x 10⁹ (OD₇₅₀ of 1.0 = 8 x 10⁸ cells/ml,OD₇₅₀=2.7A.) Table 1.2 Toxicity of LLP29 to Culex quinquefasciatus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	30	30	100
2	30	30	100
3	30	30	100
Control	30	0	0

Note. Thenconcentration of LLP29 is 2.16 x 10⁹ (OD₇₅₀ of 1.0 = 8 x 10⁸ cells/ml,OD₇₅₀=2.7A) Table 2.1 Toxicity of Chlamydomonas reinhardtii(Cry4Aa+2A+Cyt1) toA. albopictus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	100	93	93%
2	100	89	89%
3	100	91	91%
Control 1 Control 2	100	0	0

Note.1. The OD₇₅₀of Chlamydomonas reinhardtii is 0.559 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629) 3. The Control 2 group is TAP medium Table 2.2 Toxicity of Chlamydomonas reinhardtii(Cry4Aa+2A+Cyt2) toA. albopictus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	100	89	89%
2	100	90	90%
3	100	88	88%
Control 1 Control 2	100	0	0

Note1. The OD₇₅₀of Chlamydomonas reinhardtii is 0.559 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629) 3. The Control 2 group is TAP medium Table 2.3 Toxicity of Chlamydomonas reinhardtii(Cry10Aa+2A+Cyt1) toA. albopictus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	100	90	90%
2	100	92	92%
3	100	91	91%
Control 1 Control 2	100	0	0

Note1.The OD₇₅₀of Chlamydomonas reinhardtii is 0.559 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629) 3. The Control 2 group is TAP medium Table 2.4 Toxicity of Chlamydomonas reinhardtii(Cry10Aa+2A+Cyt2) toA. albopictus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	100	90	90%
2	100	90	90%
3	100	89	89%
Control 1 Control 2	100	0	0

Note1. The OD₇₅₀of Chlamydomonas reinhardtii is 0.559 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629) 3. The Control 2 group is TAP medium Table 2.5 Toxicity of Chlamydomonas reinhardtii(Cry11Aa+2A+Cyt1) toA. albopictus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	100	90	90%
2	100	95	95%
3	100	91	91%
Control 1 Control 2	100	0	0

Note1.The OD₇₅₀of Chlamydomonas reinhardtii is 0.559 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629) 3. The Control 2 group is TAP medium Table 2.5 Toxicity of Chlamydomonas reinhardtii(Cry11Aa+2A+Cyt2) toA. albopictus

Repeating group	Number of insects	Number of mortality	Corrected mortality (%)
1	100	91	91%
2	100	90	90%
3	100	89	89%
Control 1 Control 2	100	0	0

Note1. The OD₇₅₀of Chlamydomonas reinhardtii is 0.559 2. The Control 1 group is Chlamydomonas reinhardtii (notransformation, OD=0.629) 3. The Control 2 group is TAP medium

3.Reference

1.Zhang, L., Huang, E., Lin, J., Gelbič,I., Zhang, Q., Guan, Y., … Guan, X. (2010). A novel mosquitocidal strain LLP29isolated from the phylloplane of. Microbiological Research, 165(2), 133–141.doi: 10.1016/j.micres.2009.03.002

@@ Line 142: / Line 142: @@
 <p><span style="font-size:12.0pt;font-family:&quot;:,serif">&nbsp; &nbsp;&nbsp;In our project, we transform theexpression vector of <i>Cry</i> and <i>Cyt</i> toxins. Based on the toxin groups,we construct six various toxins express devices as <i>Figure1</i>. To compare the toxicities, we also tested the toxicity of <i>Bacillus thuringiensis</i> strain <i>LLP229</i>. </span><span style="font-size:12.0pt;font-family:&quot;:,serif">                               [[File:T--FAFU-CHINA--proof2.png | 825px | thumb | center ]]</span><span style="font-size:12.0pt;font-family:&quot;:,serif"></span></p>
 = <p><b><span style="font-size:12.0pt;font-family:&quot;:,serif">2.Material, Condition and Method</span></b></p> =
-<span>&nbsp; &nbsp;&nbsp;<span><p><span style="text-indent:36pt;font-size:12pt;font-family:&quot">The larvae of <i>Ae. albopictus</i> are feeding in the Key Laboratory of Biopesticideand Chemical Biology. The generation of <i>Ae.albopictus </i>is 11. The <i>LLP229</i>strain of <i>Bacillus thuringiensis</i> isprovided by Key Laboratory of Biopesticide and Chemical Biology.</span><span style="text-indent:36pt;font-family:&quot"> The concentration of </span><i style="text-indent:36pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">LLP229</span></i><span style="text-indent:36pt;font-size:12pt;font-family:&quot">is recorded in eachtable.</span></p>
+<span>&nbsp; &nbsp;&nbsp;<p><span><span style="text-indent:36pt;font-size:12pt;font-family:&quot">The larvae of <i>Ae. albopictus</i> are feeding in the Key Laboratory of Biopesticideand Chemical Biology. The generation of <i>Ae.albopictus </i>is 11. The <i>LLP229</i>strain of <i>Bacillus thuringiensis</i> isprovided by Key Laboratory of Biopesticide and Chemical Biology.</span><span style="text-indent:36pt;font-family:&quot"> The concentration of </span><i style="text-indent:36pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">LLP229</span></i><span style="text-indent:36pt;font-size:12pt;font-family:&quot">is recorded in eachtable.</span></p>
 <p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">The CC530 and FA479 strains of <i>Chlamydomonas reinhardtii </i>and TAP mediumare bought from the Aquatic Species Research Institute in Wuhan, Hubei.</span><span style="font-family:&quot;:,serif"> The concentration of </span><i><span style="font-size:12.0pt;font-family:&quot;:,serif">Chlamydomonasreinhardtii </span></i><span style="font-size:12.0pt;font-family:&quot;:,serif">is recorded ineach table.</span></p>
 <p style="text-indent:36.0pt"><span style="font-size:12.0pt;font-family:&quot;:,serif">We contrasted various toxins groupexpression vectors and transform them into <i>Chlamydomonasreinhardtii </i>as following groups: <i>Cry4Aa+2A+Cyt1,Cry4Aa+2A+Cyt2, Cry10Aa+2A+Cyt1, Cry10Aa+2A+Cyt2, Cry11Aa+2A+Cyt1,Cry11Aa+2A+Cyt2</i>.</span></p>
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Difference between revisions of "Team:FAFU-CHINA/Proof"

Revision as of 00:29, 20 October 2016

Contents

Proof

1.Introduction

2.Material, Condition and Method

<p align="center" style="text-align:center">3.Reference</p>