www.chinadairy.net zgrpgy@163.com MLSA ( 100027) : 16S rrnahsp60rpobatpdtuf (MLSA) :MLSA; ; :Q93-331 :B :1001-2230(2011)06-0046-05 Research progress of multilocus sequence analysis (MLSA) for rapid identification of Bifidobacterium species ZHAO TingCHENG Chi (China National Research Institute of Food & Fermentation Industries China Center of Industrial Culture Collection Beijing 100027 China) Abstract: This paper reviewed and evaluted the use of 16S rrnahsp60rpobatpdtuf gene part sequences as species identification tools for Bifidobacterium Strain sequences were derived from GenBank datebase Phylogenetic analysis showed that the multilocus sequence analysis(ml- SA) approach to Bifidobacterium taxonomy was possible and the discriminary of MLSA was higher than 16S rrna. Keyword: MLSA;Bifidobacterium;gene;Identification 0 1929Pribram Tissieria 1938Prevot Bifidibacterium Bifidobacterium (Bifidobacterium) 1980SKERMAN [1] 201065 (Aprroved List of Bacterial Names) 7 [3] 36 9 (B.bifidum) [4] 16S rrna 30 16S : rrna 2004Liesbeth [5] DNA-DNA (Multilocus Sequence Analysis MLSA) (B. lactis) (B. animalis subsp. lactis);2002 Sakata [6] 16S [2] rrnahsp6016s -23S ITS DNA - hsp60rpobatpdgroeltuf 1 (Bifidobacterium)1899 Tissier 1924Orla-Jensen :2011-03-31 : (No.2007BAK36B06) : (1980-) : 46 (AFLP) atpdgroel DNA (B.infantis) (B.suis) (B.longum) 3 (B.longum);2008Mattarelli [7] DNA- DNA 16S rrna hsp60 (RAPD) (DGGE) (PAGE) (B.longum) 3 : (B.longum subsp. longum) (B.longum subsp. in fantis) (B.longum
Monographs subsp. suis) 2010Jiri [8] B. actinocoloniiformeb. bohemicum Min-Soo [10] [9] B. stercoris Hidetoshi B. kashiwanohense 2 16S rrna Stackebrandt [11] 16S rrna 97% (1 )GenBank 16S rrna (Bacillus subtilis) NBRC 13719 T MEGA4.1 Kimura 1 16S rrna 1000 ( 1) : 16S rrna B.longum subsp. longum B.longum subsp. suis B.longum subsp. infantis B.breve Kimura 1000 97.6%~99.7%; groel B.animalis subsp. animalisb.animalis subsp. lactisb.choerinumb.pseudolongum 1 ; PCR 2 97.1%~99.3%; B.adolescentisB. stercorisb.ruminantiumb.dentium hsp60 (2): B.longum 97.5%~99.4%;B.bifidum subsp. longumb.longum subsp. suisb.longum subsp. in- fantis 97.0% 98.4%98.8%98.4% 94.7%; B.animalis subsp. animalisb. 3 (ML- animalis subsp. lactis SA) 97.6% 85.7%; B.breve 94.7%; B.adolescentis (2~5 ) GenBank 92.6%;B.bifidum hsp60rpobatpdtuf hsp60 92.5% (heat-shock protein)rpob RNA β (RNA polymerase β-subunit gene)atpd ATP F1 β (ATP synthase F1 beta subunit)groel (groel protein)tuf Tu(elongation factor Tu) Escherichia coli K-12 MEGA4.1 rpob (3): B.longum subsp. longumb.longum subsp. infantis 98.1% 95.6%; B.animalis subsp. 1 B.adolescentis B.bifidum B.breve B.longum subsp. infantis B.longum subsp. longum B.animalis subsp. animalis B.animalis subsp.lactis ATCC 15703 T ATCC 29521 T ATCC 15700 T ATCC 15697 T ATCC 15707 T BBMN68 ATCC 25527 T DSM 10140 T 16S rrna AP009356 AB437356 AB006658 AB116305 M58739 D86185 AB050136 hsp60 AF210319 AY004280 AF240566 AF240577 AF240578 AY004273 AY004282 GenBank rpob atpd AP009256 AY487144 EU780710 AY487145 EU780712 AY487154 CP001095 AY487150 EU780720 CP002286 EU780709 AY487152 CP001606 AY487153 groel AY488178 AY488180 tuf AY372045 AY372041 AY372046 AY372043 AY372042 AY370920 AY370919 Vol.39No.6 2011(total 247) 47
Monographs hsp60 rpob atpd groel tuf hspf1 hspr2 hspbf1 hspbr2 biff bifr bifr2 atp1 atp2 gro1 gro2 bif1 bif2 2 2 95.0%; B.adolescentisB.pseudocatenulatum (B.longum)3 PCR 5 -CAGGCGAGGATCTTGGT-3 5' -ATCGCCAAGGAGATCGAGCT -3' (hspf1-hspr2) 644 bp 5' -AAGGTGCCGCGGATCTTGTT -3' (hspbf1-hspbr2) 680 bp 5' -CCGACCATCACCAACGATGG -3' 5' -CAGAACTTGAACGTGCCACG -3' 5' TCGATCGGGCACATACGG -3' (biff-bifr) 351bp 5' CGACCACTTCGGCAACCG -3' (biff-bifr2) 523 bp 5' ACAAGATCAACCGCAAGC -3' 5' -CACCCTCGAGGTCGAAC-3' 1133bp 5' -CTGCATCTTGTGCCACTTC-3' 5'-GACCATCACCAACGATG-3' 1158bp 5'-GCTCCGGCTTGTTGGC-3' 5 -GAGTACGACTTCAACCAG-3 825bp 97.5% 96.9%;B.bifidum 96.3% atpd (4): B.longum subsp. longumb.longum subsp. suisb.longum subsp. infantis 98.7% 97.1% 96.9% 95.6%; B.animalis subsp. animalis B.animalis subsp. lactis 97.2% 94.1%; B.breve 95.6%; B. adolescentis 92.8%;B.bifidum 94.1% tuf (5): B.longum subsp. longumb.longum subsp. infantisb.bifidum 96.4% 94.0% 93.0%; B.animalis subsp. animalisb.animalis subsp. lactisb.breveb.magumb.cuniculi 96.4% 93.7% 94.3%92.8%; B.adolescentisB.ruminantium hsp60 97.6% 16S rrnahsp60rpobatpdtuf hsp60rpobatpdtuf 16S rrna 2 (B.longum) (B.longum subsp. infantis) (B. longum subsp. longum)2 RAPD DGGE (ribotyp- [7] ing) 2002Sakata 3 rpob [6] RAPD animalisb.animalis subsp. lactis (B.longum)3 97.5% 2002Requena [12] (B.longum)3 94.3%; B.breve DGGE 2006Sakata [13] 48 ribotyping (B.longum)3
Monographs 4 atpd Byoung [17] rpob rpob 84.1%~99.0% [18] hsp60 hsp60 Ventura [19] tuf 16S rrna ;Sheu [20] tuf PCR 16S rrna Zeigler [21] DNA Richter [22] ANI(Average Nucleotide Identity) DNA ANI 4 MLSA Naser [15] MLSA (Enterococcus) [14] Naser phesrpoa MLSA (B.adolescen- 16S rrna tis) (B.bifidum) (B. breve)3 Lactobacillus amylophilus LMG 11400NRRL B-4435 Lactobacillus amylotrophicus McTaggart [16] gyrb16s rrnaseca1hsp65rpob 5 5 5 tuf 3 MLSA 3 (B.longum)3 / D- B. longum subsp. longum + + + - B.longum subsp. infantis - - + + 21 (IDF) B.longum subsp. suis + - - - IDF377 110 (Bifidobacterium pseudolongum) (EFSA) (QPS) 74 MLSA API 20 A (B. infantis) (B. lactis) (L. lactis subsp. diacetylactis) MLSA Vol.39No.6 2011(total 247) 49
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