Difference between revisions of "Part:BBa K1864000"
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__NOTOC__
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<partinfo>BBa_K1864000 short</partinfo>
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Our part is CSBV's RdRP,RdRP gene cloned into the L4440 vector,and transformed into E.coli strain HT115 to express dsRdRP.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1864000 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K1864000 parameters</partinfo>
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<h3>Application </h3>
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<h3>Group:  FAFU-CHINA </h3>
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<h3>Author:  Ruicheng Dai & Changlong Lu </h3>
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<br>
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<h3>Summary: The control efficiency of dsRdRp in Chinese Scabrood Virus（CSBV）</h3>
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<h3>Design</h3>
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In our project,we have been aiming to control CSBV through silence of CSBV’s RdRp (RNA-dependent RNA polymerase) gene by using RNA-interference technology. We built recombinant L4440 plasmids which contained dual T7 promoter and we transformed it to HT115 strain.
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After the enlage cultivation of HT115, we extracted dsRdRp from bacteria. And we added different concentrations into the honeybees' food.
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Meanwhile,We noticed that the induction of IPTG could improve the expression of dsRdRp. Therefore,we also tested the inductive effect of IPTG.
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<br>
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<h3>The control efficiency of dsRdRp in Chinese Scabrood Virus</h3>
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In apiculture, scientists regard the number of sealed brood as an important index which means normal developing embryo.Therefore, we gathered datas about the effects in different dsRdRp concentrations and administration time.
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[[File:FAFU-CHIAN_Parts_information_(3).png.png|300px|thumb|left|Figure 1.Concentration of dsRdRp differs under different concentration of IPTG. The concentration of dsRdRp continues to increase till the concentration of IPTG reaches to 0.4 mmol/L]]
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<h3 >The inductive effect of different time length</h3>
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We tested dsRNA concentration with the induction time length from 0 to 9 hours. And we tested three group which involved concentration.
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[[File:FAFU-CHINA PART (2).png|300px|thumb|left|Figure 2.Concentration of dsRdRp changes with three different concentration of IPTG over time. Induced by IPTG about 4 hours, the concentration of dsRdRp appears to decrease or remain unchanged]]
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Revision as of 12:26, 15 September 2015

CSBV-RdRP

Our part is CSBV's RdRP,RdRP gene cloned into the L4440 vector,and transformed into E.coli strain HT115 to express dsRdRP.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 837
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 110
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


CSBV-RdRP

Our part is CSBV's RdRP,RdRP gene cloned into the L4440 vector,and transformed into E.coli strain HT115 to express dsRdRP.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 837
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 110
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Application

Group: FAFU-CHINA

Author: Ruicheng Dai & Changlong Lu


Summary: The control efficiency of dsRdRp in Chinese Scabrood Virus（CSBV）

Design

In our project,we have been aiming to control CSBV through silence of CSBV’s RdRp (RNA-dependent RNA polymerase) gene by using RNA-interference technology. We built recombinant L4440 plasmids which contained dual T7 promoter and we transformed it to HT115 strain. After the enlage cultivation of HT115, we extracted dsRdRp from bacteria. And we added different concentrations into the honeybees' food. Meanwhile,We noticed that the induction of IPTG could improve the expression of dsRdRp. Therefore,we also tested the inductive effect of IPTG.



The control efficiency of dsRdRp in Chinese Scabrood Virus

In apiculture, scientists regard the number of sealed brood as an important index which means normal developing embryo.Therefore, we gathered datas about the effects in different dsRdRp concentrations and administration time.

Figure 1.Concentration of dsRdRp differs under different concentration of IPTG. The concentration of dsRdRp continues to increase till the concentration of IPTG reaches to 0.4 mmol/L




















The inductive effect of different time length

We tested dsRNA concentration with the induction time length from 0 to 9 hours. And we tested three group which involved concentration.

Figure 2.Concentration of dsRdRp changes with three different concentration of IPTG over time. Induced by IPTG about 4 hours, the concentration of dsRdRp appears to decrease or remain unchanged