生物信息学教学资料：引物设计

1、引物设计生科1201班周宇筝3702120110现在要对h9n2病毒的基因设计引物。一.利用ncbi查找该病毒的全部基因1 进入ncbi的genebank界面。display settings: © tabular 20 per page. sort by relevancesond to:2输入病毒的名字h9n2,查找相关的基因。gene sources genomiccategoriesresults: 1 to 20 of 22«first < prg page 1 ot2 next> lastalternately spliced_annotated g

2、ns protein-codingname/gene id descriptionlocationaliases hahemagglutinin influenza a virusnc.004908.1 (32 1714)fluavahhh9n2s4gp1sequence contentid： 1460996(a/hong kong/10799(h9n2)ccds ensembl refseq ns1 10:1460995nonstructural protein 1 influenza a virus (a/hong kong/i073/99(h9n2)nc.004906 1 (27 719

3、)fluavahhh9n2s8gp2refseqgene m2 id: 1461002matnx protein 2 influenza a vjojs (among kong/10739(h9n2)nc.004907.1 (33.1014)fluavahhh9n2s7gp1status current onlyu eb2id: 1460998polymerase pb2 influenza a virus (miong kong/10739(h9n2)nc_004910.1 (28.2307)fluavahhh9n2s1gp1chromosome locations moren oaid：

4、1460997neuraminidase influenza a wes (a/hong kong/10739(h9n2)nc.004909 1 (1 1404)fluavahhh9n2s6gp1clear all np id: 4036076nucleoprotein unflwnza a virus (a/hong kong/10739(h9n2)nc_004905 2 (40 1536)fluavahhh9n2s5gp1show additional filters nep id: 4036374nonstructural protein 2 influenza a vjojs (a/h

5、ong kong/i073/99(h9n2)nc.004906 1 (27. 864)fluavahhh9n2s8gp1ns2 ml id 1461001matrix protein 1 influenza a virus (a/hong kong/10739(h9n2)nc.004907.1 (33 791)fluavahhh9m2s7gp2n凶広id： 13229457pa-x protein influenza a virus (a/hong kong/10739(h9n2)nc.004912 1 (21 780)fluavahhh9n2s3gp23以该病毒的一个基因h9n2ha为例，进

6、行引物设计atggagacagtatcactaataactatactactagtagcaacagtaagcaatgcagataaaatctgcatcggctatcaatcaacaaactccacagaaactgtggacacactaacagaaaacaatgtccctgtgacacatgccaaagaactgctccacacagagcataatgggatgctgtgtgcaacaagcttgggacaacctcttattttggacacctgc acaattgaagggctaatctatggcaatccttcttgtgatctatcgctggaaggaagagaatggtcctatatcgtc

7、gagag accatcagctgttcacggattgtgttaccccgggaacgtagaaaatctagaagagctaaggtcactttttagttctgcta ggtcttatcaaagaatccagattttcccagacacaatctggaatgtgtcttacgatgggacaagcacagcatgctcaggttcattctacagaagcatgagatggttgactcgaaagaacggcgattaccctatccaagacgcccaatacacaaataatcaagggaagaacattcttttcatgtggggcataaatcacccacccaccgatacta

8、cacagagagatctgtacacgagaaccgacacaacaacgagtgtggcaacagaagaaataaataggatcttcaaaccattgataggaccaaggcctcttgtcaacgg tttgatgggaagaattgattattattggtctgtattgaaaccgggtcaaacactgcgaataaaatctgatgggaatctaatagctccatggtatggacacattctttcaggagagagccacggaagaattctgaagactgatttaaaaaggggtagctg cacagtgcaatgtcagacagagaaaggtggctt

9、aaacacaacactgccattccaaaatgtaagtaggtatgcatttggaaa ctgctcaaaatacattggcataaagagtctcaaacttgcagttggtctgaggaatgtgccttctagatctagtagaggactattcggggccatagcagggtttatagagggaggttggtcaggactagttgctggttggtatgggttccagcattcaaatg accaaggggttggtatggcagcagatagagactcaacccaaaaggcaattgataaaataacatccaaagtgaataatata gtcgacaa

10、aatgaacaagcagtatgaaatcattgatcatgaattcagtgaggtagaaactagacttaacatgatcaataat aagattgatgatcaaatccaggatatatgggcatataatgcagaattgctagttctgcttgaaaaccagaaaacactcg atgagcatgacgcgaatgtaaacaatctatataataaagtaaagagggcgttgggttccaatgcggtggaagatgggaa aggatgtttcgagctataccacaaatgtgatgacca/ytgcatggagacaattcggaacgggacct

11、acaacagaaggaag tatcaagaggagtcaaaattagaaagacagaaaatagagggggtcaagctggaatctgaaggaacttacaaaatcctcac catttattcgactgttgcctcatctcttgtgattgcaatggggtttgctgccttcttgttctgggccatgtccaatgggtcttgcag/ytgcaacatttgtatataa用primers软件为这些基因设计引物。1.首先安装一个primers软件2点击file下拉菜单，选择new,然后选择dna sequencefile edit view function translate window helpnewopeninsert sequencesavesave asprefere ncesdegenerate basesprintquitctrl+sctrurqrlfb3.将已经查到的基因序列复制到文本框中。4.因为最终得到的基因其目的是插入到质粒中，因此设计引物前需要 进行酶切位点分析，所以要点击enzyme按钮，选择add添加口标酶, 然后点击0k进彳亍分析。5