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pBS423 delta repA

Bifidobacterium longum–E. coli shuttle vector.

Catalog number RDB15973
Resource name pBS423 delta repA
Clone info. Bifidobacterium longum–E. coli shuttle vector. Conditional replication (not to replicate) for the first crossover event in B. longum 105-A. Please note that pBS423 delta repA is hard to be replicated in B. longum 105-A strain. Please refer Genbank accession AB687999 for information of the original pBS423.
Comment The depositor uses spectinomycin at a concentration of 75 microgram/mL for selection of transformants in agar media. B. longum 105-A (JCM 31944) is available from JCM as a host strain.
Vector backbone pBS423 delta repA (Plasmid)
Size of vector backbone 4.4 kb
Selectable markers SpR
Depositor|Developer Fukiya, Satoru | Kano, Yasunobu |
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External Database

            Reference sequence
              
             
            Sequence (full) RDB15973zzt01.seq compiled by us

            Distribution information

            Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
            Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested. (Hirayama et al., Appl. Environ. Microbiol., 78(14):4984-4994 (2012)). The availability of the BIOLOGICAL RESOURCE is limited to a not-for-profit academic purpose.
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            提供条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する(Hirayama et al., Appl. Environ. Microbiol., 78(14):4984-4994 (2012))。本リソースは,非営利の学術研究に限って提供する。
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            RDB15973 pBS423 delta repA DNA solution

            Please review the QC test results indicated by check icon below as well as clone information before placing your order.

            How to cite this biological resource

            Materials & Methods section:

            The pBS423 delta repA was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15973).

            Reference section:

            Hirayama, Y., Sakanaka, M., Fukuma, H., Murayama, H., Kano, Y., Fukiya, S., Yokota, A., Development of a double-crossover markerless gene deletion system in Bifidobacterium longum: functional analysis of the α-galactosidase gene for raffinose assimilation. Appl. Environ. Microbiol. 78 (14): 4984-4994 (2012). PMID 22582061. [link to RRC of NBRP]

            Further references such as user reports and related articles (go to bottom)


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            Featured content

            Featured content Bacteria host strains (en) (English text)
            Featured content Bacteria host strains (ja) (Japanese text)

            QC test results

            RIKEN BRC has sequenced portions of this material for quality test.
            Please review the QC test results indicated by check icon as well as clone information before placing your order.

            Test sheet RDB15973_A8Fbp1-4.pdf check

            Nucleotide sequence of a portion of this resource (if available).

            Primer: pBS423_seq3_4 (Pr1260)
            Region: insert mid(delta repA,rep B 5')
            Sequence file: RDB15973_A8Fbe.seq check
            >15973_15973_B2LdF_M2_pBS423_seq3_4_D01_04_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNNGCTTCNC TGGGCTGANC NTCTGACTTG GTTCGGCGCA
               61 CGCTTGTGCA ATGCACAAAT GTGCAATAGG ATGTGAGCTA TGAGCAACGA ATATGTGCAG
              121 GGGACGCTGG AGCTGACGCG GGCGTTCGAC GGCTGGTGGC TTCCCCAGCG TCCCCTGTGC
              181 TGCGACGATG ACTACTCCCA GCTGGTGCGC CGGAGCCGCA TCGACGCGCT CAGATGCAAG
              241 CACATCGAGG CGAATCCGGC AGCGCAGGTG AACATGCTCG TTGTTAACGA CACCGCCAAG
              301 AGCATCCACA AGTGGATCAC CACGCGAAGC CGCATGTGGC GAGACGGTGC CGTCGCCAAC
              361 GCCGCCACGT TCGTCGCCAT CCAATCCGCA CGCGGAAAGA AATCGGGCGA GGCCAGACAA
              421 AACGCATTTG AAGAGAAGTT CGCGCAGTAC GCGCAGGAGG TGCTTGGACA ATGACCATCC
              481 AGACAATTCG CAAGAAGCGC CCATTGCCCG CCAAGGAGTT AGCCGAGGCC TATGGGGTGT
              541 CCGTGCGCAC GATCAAGTAC TGGAATTCGC AAACGCGCGA GGACTGGATA GACGAGCAGG
              601 CAACGTTGCG CGAATCCATC CNCGCCTACC ACGACGATGA CNGNNNNTCG TGGAGCCAGA
              661 CCGCCGAACA CTTCAACATG ACCCAAGGTG CCGTGCGCCN ANNNNCCTAC CGCGCNNNNN
              721 NGGAACGCGA GGCCGAGGCA AAAGCCGCAA GGNCGGANTA GCAAGTGACG TGAACNTATA
            781 TACNNTTCAC TCNNNNNNNN NN
            //
            Primer: pTBR101CM_seq3 (Pr0560)
            Region: insert mid(pTB4_p1)
            Sequence file: RDB15973_A8Fbf.seq check
            >D04829A2-14_A8Fb_pTBR101CM_seq3_B01_02_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNNC GGNCTGGNNG NTCCGCCGAN CNTGGGGCAT GGGTCTTCCC
               61 GGCAAGCCGA TTCGCACCGC GAATCGAGTG ATTGTAGCTA CAATCACGTC GCTTGCTACT
              121 CCGGCCTTCG GCACTCGGGT TGTTGCTCCT GCGCCGAAGG GCGAGGGAGG CTGCGGGCTC
              181 GGTTCATGTA CATGAACCGT CTAGCTTGCT TACCTTCGAT TTGATGGACA TCTCCATGTG
              241 GATGTCCATG GAGATGTCCA TGGAGATGTC CATGGAGATG TCCATGGAGA TGTCCATGGA
              301 GATGTCCATG TGGGTGTCCA TGTGGGTGTC CATGGATATA TCCATCTGAG CGCATAGGAA
              361 AACGCATGAG GGTGGATGAT CGTTCGGGGT GATGCCTTCT TCCACGAACG GGAAGGGGCT
              421 CTTCCTTTTT CTGGAATGTC ATCTTCCAGC GCCCTTCGCC TTGAGCTCCT TGAGCAGCAT
              481 CGTCGCGTTG TCGCGGATGC TCGCGTCCGT GAGCGGGGAG AGCTCGTCCG CATAGGAGCG
              541 GCAGAGGAGC GCGTACACCC GTTCCCGCAC CCAGGCGCTG CGCGCGATGC CCAGCTTGAA
              601 CGCGGCCCGG CACAGCATCT CGTATTCGTC GCCGGTGAGC AGGACATTGA GCTGGCGTCA
              661 AAAGTTGATT AGTCATGCCG TGTATATACG TCCGGGGTGT TTTCGGCCAG ATTCCAACAG
              721 TTTTGGGCCA CTCCGGGAAN GAACAGCCGA CGCTTCGCTG GGCTGAACCA TCTGACTTGG
              781 TTCGGCGCAC GCTTGTGCAA TGCACAAATG TGCAATAGGA TGTGAGCTAT GANCAACGAA
              841 TATGTGCAGG GGNNGCTGGA NCTGACGCNG GCGTTCGACG GCNGGNGGCT NNCCCAGCGN
              901 CCCCNNNGCT GCGNCGATGN NTACTCCNAG CNGGNGCNCC NGANCCNNNT CGACGCGCTN
            961 NNNTGNNNGC ACNTCNNNNN NNANTCCNGN NNNNNNNNN
            //
            Primer: ColE1ori_F (Pr0393)
            Region: ori,insert (pTB4_p3 5')
            Sequence file: RDB15973_A8Fbg.seq check
            >15973_15973_B2L9F_ColE1ori_F_F01_06_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNNN NNCNNNTCTG NNTTGAGCGT CGATTTTTGT GATGCTCGTC
               61 AGGGGGGCGG AGCCTATGGA AAAACGCCAG CAACGCGGCC TTTTTACGGT CCCTGGCCTT
              121 TTGCTGGCCT TTTGCTCAGA ATTCGAGCTC GGTACCCGGG GATCCAGGTG CTTGACAGGG
              181 TGATGGAGCG TCACCCAGAA CTGTCCGAGA AGGACGTGGT GACCGCGTTC CGCAGCGTCA
              241 TGGTGGACGC GGAGCGCGAG TCCGGCGCGT GGATGGCGAT AGGGCTCGAC GGGCGCGGCA
            301 GGAACGTCGA GATGCTGTAC AGGGTCG
            //
            Primer: pBS423_seq3 (Pr0559)
            Region: insert mid(delta repA 3',repB,pTB4_p2 3')
            Sequence file: RDB15973_A8Fbh.seq check
            >D04829A2-14_A8Fb_pBS423_seq3_A01_01_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNNG CTCGTTGTTN NNGACNCCGC CNAGAGCATC CACAAGTGGA
               61 TCACCACGCG AAGCCGCATG TGGCGAGACG GTGCCGTCGC CAACGCCGCC ACGTTCGTCG
              121 CCATCCAATC CGCACGCGGA AAGAAATCGG GCGAGGCCAG ACAAAACGCA TTTGAAGAGA
              181 AGTTCGCGCA GTACGCGCAG GAGGTGCTTG GACAATGACC ATCCAGACAA TTCGCAAGAA
              241 GCGCCCATTG CCCGCCAAGG AGTTAGCCGA GGCCTATGGG GTGTCCGTGC GCACGATCAA
              301 GTACTGGAAT TCGCAAACGC GCGAGGACTG GATAGACGAG CAGGCAACGT TGCGCGAATC
              361 CATCCGCGCC TACCACGACG ATGACGGCCA CTCGTGGAGC CAGACCGCCG AACACTTCAA
              421 CATGACCCAA GGTGCCGTGC GCCAACGCGC CTACCGCGCC CGCAAGGAAC GCGAGGCCGA
              481 GGCAAAAGCC GCAAGGCCGG AGTAGCAAGT GACGTGAACG TATATACGTT CACTCGATTC
              541 GCGGTGCGAA TCGGCTTGCC GGGAAGACCC ATGCCCCATA GTCGGCAGAG CCTCCAGACC
              601 GCTCACGCGG CATCGTGCCA CGACTGTGGC ACATCACGTA ACACCCGCAC ATCCTCACGG
              661 TGGTGCGCGG GTGTTACGTG ATTCGAACCA TCGGCCTGAC AGGTCTCAGA TCGCTCCAGC
              721 GAGCTTACGG CGCATGTACT CGCTGCGGCT GATGTGGTAC CGCCTGGCCA TCGCGGTCAG
              781 ANCGTCCAGC GTGCTCTTGG GCATGCGGAC GGACACGCTG ACCATCTGCT CGNCGGGCTC
              841 GGCGANATGG ANCCCGTAGT ACACGCGCCC CGTCAGGTCG TCCGGGNNNN TCTCGGANTC
              901 NGCCATCCGC NCATCCTCCA NCACCNGCTC CTCNGNCAGC CCNANCAGCG CCNTCAANNG
            961 NNNGNCNNNT TCGCNGAAAN TCNCNNATAN NNNNNNNNN
            //
            Primer: pTBR101CM_seq7 (Pr0561)
            Region: insert mid(repB 3',pTB4_p2,pTB4_p3 3')
            Sequence file: RDB15973_A8Fbi.seq check
            >D04829A2-14_A8Fb_pTBR101CM_seq7_C01_03_ABI08.ab1
                1 NNNNNNNNNN NNNNNNNNNC CGCGCCTACN NCNACNNNGA CGGCCNCTCG TGGNGCCAGA
               61 CCGCCGAACA CTTCAACATG ACCCAAGGTG CCGTGCGCCA ACGCGCCTAC CGCGCCCGCA
              121 AGGAACGCGA GGCCGAGGCA AAAGCCGCAA GGCCGGAGTA GCAAGTGACG TGAACGTATA
              181 TACGTTCACT CGATTCGCGG TGCGAATCGG CTTGCCGGGA AGACCCATGC CCCATAGTCG
              241 GCAGAGCCTC CAGACCGCTC ACGCGGCATC GTGCCACGAC TGTGGCACAT CACGTAACAC
              301 CCGCACATCC TCACGGTGGT GCGCGGGTGT TACGTGATTC GAACCATCGG CCTGACAGGT
              361 CTCAGATCGC TCCAGCGAGC TTACGGCGCA TGTACTCGCT GCGGCTGATG TGGTACCGCC
              421 TGGCCATCGC GGTCAGACCG TCCAGCGTGC TCTTGGGCAT GCGGACGGAC ACGCTGACCA
              481 TCTGCTCGTC GGGCTCGGCG AGATGGAGCC CGTAGTACAC GCGCCCCGTC AGGTCGTCCG
              541 GGACAGTCTC GGACTCGGCC ATCCGCTCAT CCTCCAGCAC CTGCTCCTCG GTCAGCCCGA
              601 CCAGCGCCAT CAACTGCCTG TCGCGTTCGC TGAAATCACT CATAGCGTCC TCCCATCGCC
              661 GCGCAGACGG TCTATCTCGC GCCTGAACTT CTTGGTCGGC GGCGTGAACG CGTGGTAGAT
              721 CACCACGAGG TCGCCGACGA CCCTGTACAG CATCTCGACG TTCCTGCCGC GCCCGTCGAG
              781 CCCTATCGCC ATCCACGCGC CGGACTCGCG CTCCNCGTCC ACCATGACGC TGCGGAACGC
              841 GGTCACCACG TCCTTCTCGG ACAGTTCTGG GGTGANGCNC CATCACCCNN NCAAGCACCN
              901 NGATCCCNGG GNNNCNNNCT NNNNNTCTGA NNNAAAGGNC AGCNAAANGN CAGGGANCGN
            961 NAAAANNNNC NNNTNNNNNN NNTTTTTTNC ANNNNNNNNN N
            //

            Please visit Sequencing and PCR primers for primer information.


            References

            Original, user report and related articles

            original Hirayama, Y., Development of a double-crossover markerless gene deletion system in Bifidobacterium longum: functional analysis of the α-galactosidase gene for raffinose assimilation. Appl. Environ. Microbiol. 78 (14): 4984-4994 (2012). PMID 22582061. [link to RRC of NBRP]
            reference O'Connell Motherway, M., Carbohydrate Syntrophy enhances the establishment of Bifidobacterium breve UCC2003 in the neonatal gut. Sci. Rep. 8 (1): 10627 (2018). PMID 30006512.
            reference Nishiyama, K., Bifidobacterium bifidum Extracellular Sialidase Enhances Adhesion to the Mucosal Surface and Supports Carbohydrate Assimilation. MBio 8 (5). pii: e00928-17 (2017). PMID 28974612.
            reference Fukiya, S., Development of practical gene-mutagenesis systems in bifidobacteria. (Japanese text) Seibutsu-kogaku 94 (3): 110-116 (2016).

            2024.03.26

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