hela细胞转染实验

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1、1•真核表达载体构建csrB:碱基数：369bp(Hfq依赖性)2883155..2883523gctagcGTCGACAGGGAGTCAGACAACGAAGTGAACATCAGGATGATGACACTTCTGCAGGACACACCAGGATGGTGTTTCAGGGAAAGGCTTCTGGATGAAGCGAAGAGGATGACGCAGGACGCGTTAAAGGACACCTCCAGGATGGAGAATGAGAACCGGTCAGGATGATTCGGTGGGTCAGGAAGGCCAGGGACACTTCAGGATGAAGTATCACATCGGG

2、GTGGTGTGAGCAGGAAGCAATAGTTCAGGATGAACGATTGGCCGCAAGGCCAGAGGAAAAGTTGTCAAGGATGAGCAGGGAGCAACAAAAGTAGCTGGAATGCTGCGAAACGAACCGGGAGCACTGTGAATACAGTGCTCCCTTTTTTTATTtgtaca引物：上游csrB-F：gctagcGTCGACAGGGAGTCAGACAANhel酶切位点下游csrB-R：tgtacaAATAAAAAAAGGGAGCACTGBsrGI酶切位点DetailedprimerreportsPr

3、imerpair4Sequence(5'->3*)LengthTmGC%SelfcomplementaritySelf3*complementarityForwardprimerGCTAGCGTCGACAGGGAGTCAGACAA2667.8757696.001.00ReverseprimerACATGTAATAAAAAAAGGGAGCACTG2658.6234626.003.00Productsontargettemplates>NZNEOR010000011Shiaellaflexneri2astr301Scaffold1.whol

4、eoenorneshotqunsequenceproductlength=381Fomrdpri*er1CaACCCTCGACKCGACTCACACAA26Teaplate964141ATCU964116Revtrs*priatr1AaTCTMTAAAAMACGGACCACTC26Teiplate963761.GCA.C963786>NC_004337.2Shigellafteinen2astr.301chromosome.completegenomeproducts331Forwardprlxr1GCTAGCCTCGACAGCCAGI

5、CAGACAA26Isolate2S93529ATCn2383504Reversepriaer1ACATGTAATAmUAGGGAGCACTC26Template2&93149・(XXG2383)74MicF:碱基数：93bp2334487..233457SGCTATCATCATTAACTTTATTTATTACCGTCATTCATTTCTGAATGTCTGTTTACCCCTATTTCAACCGGATGCCTCGCATTCGGTTTTTTTgctagcAAGATGTTTCATTTATCGCCATAGATGTTTCAAAATGTAAATAC

6、AAGGGAACTTTTTAAGATTATTGCGGAATGGCGAAATAAGCACCTAACATCAAGCAATAATAATTCAAGGTTAAAATCAATAACTTATTCTTAAGTATTTGACAGCACTGAATGTCAAAACAAAACCTTCACTCGCAACTAGAATAACTCCCGCTATCATCATTAACTTTATTTATTACCGTCATTCATTTCTGAATGTCTGTTTACCCCTATTTCAACCGGATGCCTCGCATTCGGTTTTTTTACCCTTCTTTACACACTTTTCATTATT

7、CTGTGCTACCACAGAAAAACTATAACGCTTGTTAACTATTTCACAAATAATTAACATCAGCATAATTTCCAGCAATCTTTGTTTATTTGCAATTAATTTCGTTGGGCTTTTTGTAGGTTATTTGTACAGC引物：上游MicF-F：gctagcAAGATGTTTCATTTATCGCCNhel酶切位点下游MicF-R：tgtacaGCTGTACAAATAACCTACABsrGI酶切位点DetailedprimerreportsPrimerpair1Sequence(5*->3*)Leng

8、thTmGC%SelfcomplementaritySelf3*complementarityForv/ardprimerGCTAGCAAGATGTTTCATTTATCGCC2661.2742.316.00