pHB201
pHB201
编号 | 载体名称 |
北京华越洋VECT75425 | pHB201 |
pHB201载体基本信息:
启动子: | T7,T3 |
复制子: | ori |
终止子: | T1 |
质粒分类: | 广宿主系列,枯草杆菌载体 |
质粒大小: | 6594bp |
原核抗性: | Chl |
克隆菌株: | DH5α |
培养条件: | 37℃,有氧 LB |
表达宿主: | 枯草芽孢杆菌 |
诱导方式: | IPTG诱导 |
5'测序引物: | T7:TAATACGACTCACTATAGGG |
3'测序引物: | 根据序列设计引物 |
pHB201载体图谱:
pHB201载体简介:
A genomic library of Bacillus lyticus was constructed in lambda GEM 11 vector and screened for the xylanase gene using Congo red plate assay. A 16-kb fragment containing the xylanase gene was obtained which was further subcloned using Mbo I partial digestion in an E. coli pUC 19 vector. A 1.3-kb sub-fragment was obtained which coded for a xylanase gene of Mr 23,650 Da. This fragment was sequenced and the homology was checked with known xylanases. The maximum homology was 97%, which was obtained with an endo xylanase gene from Bacillus species at the DNA level, while the translated sequence showed only one amino acid change from alanine to serine at position number 102. Expression was checked in E. coli, using the native promoter, and an extracellular activity of 5.25 U/mL was obtained. Cloning of the gene was done in Bacillus subtilis using a shuttle vector pHB 201, which resulted in increasing the basal level xylanase activity from 14.02 to 22.01 U/mL.
pHB201载体序列:
ORIGIN
1 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
61 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
121 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
181 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
241 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
301 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
361 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
421 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
481 agtcagtgag cgaggaagcg gaagaagctt actttattgt tttgccattt taaaaggttc
541 aaataaaata tttttcataa aattatcaga tgttttttaa aaaatgaaat aaaagaagaa
601 aaaaatagaa aatacactaa cccaaagaag cgcgtaatat cgagggttta agagacaata
661 agaaaaaaag attgaaaaat gacattaaat tcttgacagg gagagatagg tttgatagaa
721 tataatagtt gtcgcgagag acgacgttga agcgcgacaa gctagaccat tgaaaactga
781 ataaagaaga atgactcatg tgatgtagca atacatcaaa aatctgtcaa tcaataatga
841 aagacaagcc atcaattcgg tgacttaaat actttatttg agagtttgat cctctagagt
901 cgacctgcag gctttaacgt aggcaaagct cagggtagac tttgaatgga cagaaacatg
961 acatatctct tgaaaggatg attgtggtgg tgaaaacaga taaaatctcc tcctgaatac
1021 agtaaatcac attcaggagg agataaaatt gtttaaacaa atagacgaaa attatctgcg
1081 aaaagagcac tttcaccatt atatgacgtt aacccgatgc tcatatagct tggtgatcaa
1141 tctagacatc acgaaattgc atgcaatatt aaaagaaaaa aagctgaaag tatatcctgt
1201 gcaaatttat ttgttagcaa gagctgtgca aaaaattcct gagtttcgga tggatcaagt
1261 gaacgatgaa cttggttact gggagattct ccatcctagt tatacgattc taaataaaga
1321 aacaaagacg ttttcaagta tttggacgcc ttttgatgaa aactttgctc agttttataa
1381 aagctgtgta gccgatattg aaacatttag caaaagcagc aacctatttc cgaaacctca
1441 tatgccagaa aacatgttca atatttcaag tctaccgtgg attgatttta cttcttttaa
1501 ccttaatgta tctacagatg aagctagcgc gcaattaacc ctcactaaag ggaacaaaag
1561 ctggagctcc accgcggtgg cggccgctct agaactagtg gatcccccgg gctgcaggaa
1621 ttcgatatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccaattcgcc
1681 ctatagtgag tcgtattacg cgcgcaattc actggccgtc gttttacaac gtcgtgactg
1741 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
1801 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatta
1861 gagatcaatt ccctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc
1921 agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc
1981 acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc
2041 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
2101 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
2161 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
2221 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
2281 cgtttctaca aactcttcct gtcgtcatat ctacaagcca tccccccaca gatacggtaa
2341 actagcctcg tttttgcatc aggaaagcag ggaattgatc tgatgggtag aaccttgggc
2401 tttgcccaaa cccaccaacg gatggccgtt ggattccaac attcaatccc cccaaccccc
2461 cgcatctccg gggggctccc tcgccggttg gcgtcaaaca aacgagtttg tttgagtgct
2521 gaaagggttc gggtagcact gagcgagtca actccttaat cctccctacg gtccggttac
2581 cgttgactca cgccataccc atcgaatggc gaatggcgga tctggagctg taatataaaa
2641 accttcttca actaacgggg caggttagtg acattagaaa accgactgta aaaagtacag
2701 tcggcattat ctcatattat aaaagccagt cattaggcct atctgacaat tcctgaatag
2761 agttcataaa caatcctgca tgataaccat cacaaacaga atgatgtacc tgtaaagata
2821 gcggtaaata tattgaatta cctttattaa tgaattttcc tgctgtaata atgggtagaa
2881 ggtaattact attattattg atatttaagt taaacccagt aaatgaagtc catggaataa
2941 tagaaagaga aaaagcattt tcaggtatag gtgttttggg aaacaatttc cccgaaccat
3001 tatatttctc tacatcagaa aggtataaat cataaaactc tttgaagtca ttctttacag
3061 gagtccaaat accagagaat gttttagata caccatcaaa aattgtataa agtggctcta
3121 acttatccca ataacctaac tctccgtcgc tattgtaacc agttctaaaa gctgtatttg
3181 agtttatcac ccttgtcact aagaaaataa atgcagggta aaatttatat ccttcttgtt
3241 ttatgtttcg gtataaaaca ctaatatcaa tttctgtggt tatactaaaa gtcgtttgtt
3301 ggttcaaata atgattaaat atctcttttc tcttccaatt gtctaaatca attttattaa
3361 agttcatttg atatgcctcc taaattttta tctaaagtga atttaggagg cttacttgtc
3421 tgctttcttc attagaatca atcctttttt aaaagtcaat attactgtaa cataaatata
3481 tattttaaaa atatcccact ttatccaatt ttcgtttgtt gaactaatgg gtgctttagt
3541 tgaagaataa agaccacatt aaaaaatgtg gtcttttgtg tttttttaaa ggatttgagc
3601 gtagcgaaaa atccttttct ttcttatctt gataataagg gtaactattg ccgcgcctga
3661 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataagc cggtcttttt
3721 cgattttggc gaagccgaaa tcgggtcttt tcttatcttg atactatata gaaacaacat
3781 catttttcaa aattaggtca aagccttgtg tatcaagggt ttgatggttc tttgacaggt
3841 aaaaactcct tctgctatta ttaaggtgtc gaatcaaaat aatagaatgc tagagaacta
3901 gctcagaagg agtttttttg ttgatttatt catctgaaaa tgattatagc atcctcgaag
3961 ataaaaccgc aacaggtaaa aagcgggatt ggaaggggaa aaagagacgg acgaacctca
4021 tggcggagca ttacgaagcg ttagagagta agattggggc accttactat ggcaaaaagg
4081 ctgaaaaact aattagttgt gcagagtatc tttcgtttaa gagagacccg gagacgggca
4141 agttaaaact gtatcaagcc catttttgta aagtgaggtt atgtccgatg tgtgcgtggc
4201 gcaggtcgtt aaaaattgct tatcacaata agttgatcgt agaggaagcc aatagacagt
4261 acggctgcgg atggattttt ctcacgctga cgattcgaaa tgtaaaggga gaacggctga
4321 agccacaaat ttctgcgatg atggaaggct ttaggaaact gttccagtac aaaaaagtaa
4381 aaacttcggt tcttggattt ttcagagctt tagagattac caaaaatcat gaagaagata
4441 catatcatcc tcattttcat gtgttgatac cagtaaggaa aaattatttt gggaaaaact
4501 atattaagca ggcggagtgg acgagccttt ggaaaaaggc gatgaaattg gattacactc
4561 caattgtcga tattcgtcga gtgaaaggta aagctaagat tgacgctgaa cagattgaaa
4621 acgatgtgcg gaacgcaatg atggagcaaa aagctgttct cgaaatctct aaatatccgg
4681 ttaaggatac ggatgttgtg cgcggtaata aggtgactga agacaatctg aacacggtgc
4741 tttacttgga tgatgcgttg gcagctcgaa ggttaattgg atacggtggc attttgaagg
4801 agatacataa agagctgaat cttggtgatg cggaggacgg cgatctggtc aagattgagg
4861 aagaagatga cgaggttgca aatggtgcat ttgaggttat ggcttattgg catcctggca
4921 ttaaaaatta cataatcaaa taaaaaaagc agacctttag aaggcctgct tttttaacta
4981 acccatttgt attgtgttga aatatgtttt gtatggtgca ctctcagtac aatctgctct
5041 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
5101 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
5161 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
5221 ctatttttat aggttaatgt catgataata atggtttctt agcgattcac aaaaaatagg
5281 cacacgaaaa acaagttaag ggatgcagtt tatgcatccc ttaacttact tattaaataa
5341 tttatagcta ttgaaaagag ataagaattg ttcaaagcta atattgttta aatcgtcaat
5401 tcctgcatgt tttaaggaat tgttaaattg attttttgta aatattttct tgtattcttt
5461 gttaacccat ttcataacga aataattata cttttgttta tctttgtgtg atattcttga
5521 tttttttcta cttaatctga taagtgagct attcacttta ggtttaggat gaaaatattc
5581 tcttggaacc atacttaata tagaaatatc aacttctgcc attaaaagta atgccaatga
5641 gcgttttgta tttaataatc ttttagcaaa cccgtattcc acgattaaat aaatctcatt
5701 agctatacta tcaaaaacaa ttttgcgtat tatatccgta cttatgttat aaggtatatt
5761 accatatatt ttataggatt ggtttttagg aaatttaaac tgcaatatat ccttgtttaa
5821 aacttggaaa ttatcgtgat caacaagttt attttctgta gttttgcata atttatggtc
5881 tatttcaatg gcagttacga aattacacct ctttactaat tcaagggtaa aatggccttt
5941 tcctgagccg atttcaaaga tattatcatg ttcatttaat cttatatttg tcattatttt
6001 atctatatta tgttttgaag taataaagtt ttgactgtgt tttatatttt tctcgttcat
6061 tataaccctc tttaatttgg ttatatgaat tttgcttatt aacgattcat tataaccact
6121 tattttttgt ttggttgata atgaactgtg ctgattacaa aaatactaaa aatgcccata
6181 ttttttcctc cttataaaat tagtataatt atagcacgaa aaggatctag gtgaagatcc
6241 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
6301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
6361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
6421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
6481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
6541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt ctta
//
pHB201载体其他相关的枯草杆菌载体: