卡氏肺孢子虫PCR引物设计

卡氏肺孢子虫PCR引物 major surface antigen MSG1 [Pneumocystis carinii] GenBank: D17438.1 GAACTCTGCACAACATGGGGCCAACATTGTCACCAACTTGTGGAGAATTGTCCAGATCA GTTGAAAAAAGAGAATGGCAATAATCATAACTGTGATGAACTCGATGAAAAATGCAGTG ATACCTTTAAAAGGTTGAAATTAGAGGAGGAGCTGACTCATCTGTTGAAAGGCAGTTTA AAGAAGGATGATCAATGTACAGAAGCATTAGGAAAGAATTGCACTGACTTGCAAAAGAA TGAAACATTCAAAATTCTGCTTAGTGATTGTGAAAAAGATAAGCAAAATGTTTGCACAA AATTAGTTAAAAAAGTACAAGAGAGATGTCCTACTTTAAAAGCTGATCTGGATAAAGCG AAAGATGAGTTGACAAAGATGAAGAATGAGTACGAAAAGGCAAAAGAGGAGGCAGAA AAATCTACAAATAAAGCTAGCTTATTGCTATCAAAGTCTGGAAAAGCCGCAATGCCAAGT GGACAGGATAGCAATGGCTCTGTACCAGCACCACCAGCAGAGCCAGCACCAGGGACAC TACCACCACCACCATCAACTGACGGAACGGTAAACACGCCAGCTGGAGGATCAGGGAC ATCCGCTGGAGGATCAGGCACACCAAGTAGTGGAACGACAGACTCTGCAAAACTTGGA CTCGTTAAAAGAGCGTATGTAGAAGGAGGAGTATCAGAAGTAGAGGTAAAAGCATTTGA TGCAACGACGGTAGCATTGGAATTGTATTTGGAATTGAAAGAGGAATGCAATGCTTTAGA ACTAGATTGCGGTTTTAAAGAGGATTGTTCAGATCTTCAAGTTTGCGGAGAAATAGACAC GTTATGCGGAGGAATAAAACCATTAGAAATTAAGCCTCATCATACAGAGACAATAACAAA TAAGGTGACGGAAACGAAGACGGAAACAAAGACAGAAACAAAGACTGATGACAAGGC TGATGAGAAGACCGGTACGAAAACTGTTACAGAAACCAAGACAATAGGTGGAGGAAAA GTAACAGAAGAGTGTACAATGATACAAACAACAGATACATGGGTAACACGTACATCATT GCATACGAGTACGACAACGAGTACGTCAACGGTGACGTCGACAGTGACGTTGACGTCGA TGCGCAAGTGCAAGCC GGCTTGCACTTGCGCATCGAC 1 atgagattga tgagaaacac cttttggctt tcattgtgaa ggacaaatat aaagaagaac 61 aaaaatgcaa agaagaactc gagaaatatt gtaaagagtt gaaggaagca gataaaaatc 121 tagagaatgt ggatgataaa gttaaaggac tttgtgatga taaaaaacga gacgaaaaat 181 gcaaagacgt gaaaaaaaaa gttgaagatg aattaaaaga ttttgaagag gaacttcaaa 241 aagtattgaa taatataaaa gatgaaaatt gcgaaaaata tgaagaaaaa tgtatacttt 301 tagaagagac ggattatgat gttattaagg ataactgtgt caagttgagg gaaggatgtt 361 acgaattgaa gcgtaaaaag gtggcagagg agctcctttt gagggcgctc ggaaaggaag 421 ctaaagaaga agttaaatgt caagcagaga tgaaaaaggt ttgcccagtg ttaagccgag 481 aaagcgacga attgatgttt ttgtgccttg attcggatgg aacgtgtcaa gcgctgaaaa 541 aaaaatcaga agaagtttgc cagcttttaa aagaaaagct taaagatggc gaattaaagg 601 aaaaatgtca tgaaagactt gagaaatgtc atttttacgg agaagcgtgt gataaaacaa 661 aatgtgatga ggataaggat caatgcgaga aaaaagaaat cacatataaa gcgccagaat 721 ctgattctag tcctgtcaag ccgaagacgt cgttgttgag aagtattggg ttggatgatg 781 tgtataaaag agctgaaaaa gaaggaatta ttattggaaa atcaggagtg gatctaccaa 841 gaaagtcagg tacaaaattt ctgcaagatc tcttgctagt gttgagcaga aatgagaatg 901 ataaggatgc ggggaagaaa tgcaccgaag cgttaaaaaa atgcaaagat cctaagtatt 961 tggatcataa tttggaagag ttatgcgata aaaatgataa cgacaaatgc aaaaaaatac 1021 tagatgtaaa agaaagatgt acaaatctca aattaaaact ttatctcaaa gggttgtcta 1081 caaaatatga tgataaagaa tcagaacttt tatcgtgggg aaagcttcca acctttttta 1141 taaaaggaga gtgtgcagaa cttgagtcgg aatgtttcta tttagaaaag gcgtgtaata 1201 ataagattga tgaagcatgt caaaatgcaa gagcagcgtg ctataaaaag ggacaagaca 1261 ggatgttgaa taagttcttt caaaaggaat tgaggggaaa gcttggtctt gtaagatatc 1321 gtagcgatcc taaagaatgc aaaaaatatg tggtagaaaa ctgtacaaaa cttaaagaag 1381 ataaaagata cctttcaaaa tgtctttatc ctaaagaact atgttatggg ctttcaaatg 1441 atatttttct tcaatccaaa gagttaagtt tgcttttaga tgatcagaga gattttccat 1501 ttgaaaagga ttgtgttgaa ttgaaggaga agtgtgatga acttggtagt gattcattat 1561 tgaatttaga aaagtgtata acattgaaaa gacgctgtga atactttaag gttacagagg 1621 gatttagaaa cgtattttta gaaaaaaagg atgattcgtt aatgactcag gaaaactgta 1681 caaaggcatt gcatgagaaa tgtaatgctt tatctaggag gagaaagaat tcatttagtg 1741 tttcatgtgc tttaccagaa gaaacatgta gttatatggt attccacaca agtcaagatt 1801 gtaaatattt aaaagaaaac atcaagaatg gaggaattgt aggaaaaatt gggggagcaa 1861 atggaaatga agcaacactc gaagaactct gcacaacatg gggccaacat tgtcaccaac 1921 ttgtggagaa ttgtccagat cagttgaaaa aagagaatgg caataatcat aactgtgatg 1981 aactcgatga aaaatgcagt gataccttta aaaggttgaa attagaggag gagctgactc 2041 atctgttgaa aggcagttta aagaaggatg atcaatgtac agaagcatta ggaaagaatt 2101 gcactgactt gcaaaagaat gaaacattca aaattctgct tagtgattgt gaaaaagata 2161 agcaaaatgt ttgcacaaaa ttagttaaaa aagtacaaga gagatgtcct actttaaaag 2221 ctgatctgga taaagcgaaa gatgagttga caaagatgaa gaatgagtac gaaaaggcaa 2281 aagaggaggc agaaaaatct acaaataaag ctagcttatt gctatcaaag tctggaaaag 2341 ccgcaatgcc aagtggacag gatagcaatg gctctgtacc agcaccacca gcagagccag 2401 caccagggac actaccacca ccaccatcaa ctgacggaac ggtaaacacg ccagctggag 2461 gatcagggac atccgctgga ggatcaggca caccaagtag tggaacgaca gactctgcaa 2521 aacttggact cgttaaaaga gcgtatgtag aaggaggagt atcagaagta gaggtaaaag 2581 catttgatgc aacgacggta gcattggaat tgtatttgga attgaaagag gaatgcaatg 2641 ctttagaact agattgcggt tttaaagagg attgttcaga tcttcaagtt tgcggagaaa 2701 tagacacgtt atgcggagga ataaaaccat tagaaattaa gcctcatcat acagagacaa 2761 taacaaataa ggtgacggaa acgaagacgg aaacaaagac agaaacaaag actgatgaca 2821 aggctgatga gaagaccggt acgaaaactg ttacagaaac caagacaata ggtggaggaa 2881 aagtaacaga agagtgtaca atgatacaaa caacagatac atgggtaaca cgtacatcat 2941 tgcatacgag tacgacaacg agtacgtcaa cggtgacgtc gacagtgacg ttgacgtcga 3001 tgcgcaagtg caagcc PC-GMG1: F: GAACTCTGCACAACATGGGGCC Tm58 PC-GMG1: R: 5，GGCTTGCACTTGCGCATCGAC 3‘ Tm 58 产物长度1133个碱基 同源性分析: 1.上游引物: >dbj|D82031.1| Pneumocystis carinii gene for major surface glycoprotein MSG105, exon1-2, complete cds Length=11363 Score = 45.5 bits (22), Expect = 0.003 Identities = 22/22 (100%), Gaps = 0/22 (0%) Strand=Plus/Plus Query 2 GAACTCTGCACAACATGGGGCC 23 |||||||||||||||||||||| Sbjct 3346 GAACTCTGCACAACATGGGGCC 3367 >emb|CR730243.1| P. carinii cosmid 17D7 Length=35111 Score = 45.5 bits (22), Expect = 0.003 Identities = 22/22 (100%), Gaps = 0/22 (0%) Strand=Plus/Plus Query 2 GAACTCTGCACAACATGGGGCC 23 |||||||||||||||||||||| Sbjct 12375 GAACTCTGCACAACATGGGGCC 12396 Score = 45.5 bits (22), Expect = 0.003 Identities = 22/22 (100%), Gaps = 0/22 (0%) Strand=Plus/Plus Query 2 GAACTCTGCACAACATGGGGCC 23 |||||||||||||||||||||| Sbjct 28062 GAACTCTGCACAACATGGGGCC 28083 Score = 37.3 bits (18), Expect = 1.0 Identities = 21/22 (96%), Gaps = 0/22 (0%) Strand=Plus/Plus Query 2 GAACTCTGCACAACATGGGGCC 23 ||||| |||||||||||||||| Sbjct 24564 GAACTTTGCACAACATGGGGCC 24585 Score = 31.2 bits (15), Expect = 71 Identities = 18/19 (95%), Gaps = 0/19 (0%) Strand=Plus/Plus Query 3 AACTCTGCACAACATGGGG 21 ||||||| ||||||||||| Sbjct 9829 AACTCTGTACAACATGGGG 9847 2(下游引物: >ref|NP_829258.1| hypothetical protein CCA00389 [Chlamydophila caviae GPIC] gb|AAP05136.1| conserved hypothetical protein [Chlamydophila caviae GPIC] Length=726 GENE ID: 1218074 CCA00389 | hypothetical protein [Chlamydophila caviae GPIC] (10 or fewer PubMed links) Score = 36.7 bits (79), Expect = 0.37 Identities = 14/20 (70%), Positives = 14/20 (70%), Gaps = 3/20 (15%) Query 4 GTCGATGCGCAAGTGCAAGC 23 G CG TGCGCA G AAGC Sbjct 360 GNCGCTGCGCAEG---AAGC 376 >gb|EFN56627.1| hypothetical protein CHLNCDRAFT_144408 [Chlorella variabilis] Length=486 Score = 35.4 bits (76), Expect = 0.89 Identities = 11/13 (85%), Positives = 11/13 (85%), Gaps = 0/13 (0%) Query 2 AAGTCGATGCGCA 14 AAGTCGA GC CA Sbjct 248 AAGTCGAAGCSCA 260 Score = 23.1 bits (47), Expect = 4505 Identities = 8/9 (89%), Positives = 8/9 (89%), Gaps = 0/9 (0%) Query 10 GCGCAAGTG 18 GCG AAGTG Sbjct 391 GCGAAAGTG 399 >ref|XP_002012139.1| GI16598 [Drosophila mojavensis] gb|EDW17270.1| GI16598 [Drosophila mojavensis] Length=994 GENE ID: 6586528 Dmoj\GI16598 | GI16598 gene product from transcript GI16598-RA [Drosophila mojavensis] (10 or fewer PubMed links) Score = 32.5 bits (69), Expect = 7.0 Identities = 15/30 (50%), Positives = 15/30 (50%), Gaps = 9/30 (30%) Query 4 GTCG-------ATGCGCA--AGTGCAAGCC 24 G CG ATGCGCA G G GCC Sbjct 859 GSCGKPKPAPAATGCGCAPSGGSGGGGGCC 888