Difference between revisions of "Part:BBa K1439000:Experience"

(→‎Test1. Conjugation between E.coli HB101 and Top10)
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===Test1. Conjugation between E.coli HB101 and Top10===
 
===Test1. Conjugation between E.coli HB101 and Top10===
 
We do conjugation experiment between HB101 and Top10 to test the conjugation ability of OriTRP4 (BBa_K1439000). We got a special strain from other lab which is sensitive to Streptomycin, while Top10 has streptomycin resistance. In order to test BBa_K1439000 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance.
 
We do conjugation experiment between HB101 and Top10 to test the conjugation ability of OriTRP4 (BBa_K1439000). We got a special strain from other lab which is sensitive to Streptomycin, while Top10 has streptomycin resistance. In order to test BBa_K1439000 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance.
  
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===Applications of BBa_K1439000===
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Conjugation between E.coli HB101 and Top10
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We have test the oriTRP4 BBa_K1439000 in a conjugation system in which pSB1C3 with the help of plasmid RP4, and ''E. coli'' HB101 as donor and'' E.coil'' Top10 as recipient cells. We test the conjugative competence of oriTRP4 BBa_K1439000 by conjugate between HB101 and Top10. We use the RFP (BBa_J04450) marks the plasmids containing oriTRP4 BBa_K1439000. We get HB101 without streptomycin resistance form Zhigang Qiu. Top 10 has streptomycin resistance. After conjugation, we select recipient cells by streptomycin resistance LB medium and statistical number of red bacterial colony.
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===User Reviews===
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!    !! Conjugation (HB101 and Top10 mixed system) !! Top10(no plasmid) !! HB101(double plasmids system)
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<!-- DON'T DELETE --><partinfo>BBa_K1439000 StartReviews</partinfo>
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<!-- Template for a user review
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| Streptomycin || White colony || White colony || No colony
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| chloramphenicol || Red colony || No colony || Red colony
 
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| chloramphenicol & streptomycin || Red colony || No colony || No colony
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<partinfo>BBa_K1439000 AddReview number</partinfo>
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|}
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<I>Username</I>
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Table.1 We constructed a new part BBa_K1439001 to test the conjugation result. BBa_K1439001 is used in conjugation experiments below.
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Enter the review inofrmation here.
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Test
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Conjugate between ''E.coil'' HB101 and ''E.coil'' Top10.
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We designed a orthogonal experiment, it is showed in the table. The first column shows the strains used in coating, and the first row shows the antibiotics added in medium. Besides, the table shows the result ideally. The second row proves that HB101, plasmid RP4 and the backbone pSB1C3 don’t contain streptomycin resistant gene. The third row proves that Top10 is sensitive to chloramphenicol. And the forth row proves that Top10 can receive the mini plasmid after conjugation.
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=Result =

Revision as of 12:56, 16 October 2014

Test1. Conjugation between E.coli HB101 and Top10

We do conjugation experiment between HB101 and Top10 to test the conjugation ability of OriTRP4 (BBa_K1439000). We got a special strain from other lab which is sensitive to Streptomycin, while Top10 has streptomycin resistance. In order to test BBa_K1439000 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance.

Conjugation between E.coli HB101 and Top10

Conjugation (HB101 and Top10 mixed system) Top10(no plasmid) HB101(double plasmids system)
Streptomycin White colony White colony No colony
chloramphenicol Red colony No colony Red colony
chloramphenicol & streptomycin Red colony No colony No colony

Table.1 We constructed a new part BBa_K1439001 to test the conjugation result. BBa_K1439001 is used in conjugation experiments below.

We designed a orthogonal experiment, it is showed in the table. The first column shows the strains used in coating, and the first row shows the antibiotics added in medium. Besides, the table shows the result ideally. The second row proves that HB101, plasmid RP4 and the backbone pSB1C3 don’t contain streptomycin resistant gene. The third row proves that Top10 is sensitive to chloramphenicol. And the forth row proves that Top10 can receive the mini plasmid after conjugation.

Result