1、人肌红蛋白基因外显子一的生物信息学分析一 Introduction：Myoglobin, the article is composed of a peptide chains and a binding protein of heme prosthetic group, is within the muscle oxygen storage protein, its oxygen saturation of the elliptic curve.Myoglobin is the main protein of skeletal muscle and cardiac muscle, when
2、muscle damage, leakage to the circulating in the blood in muscle tissue, increase the serum concentration of myoglobin, the index is used to determine whether muscle damage.二 Purpose：对人肌红蛋白基因外显子一基因序列使用 “Bioedit 和 DNAMAN、anthe5.0”及相关网站进行 GC 含量统计、电子酶切、引物设计、基因在人类基因组中的定位、电子克隆及蛋白在细胞内的定位、氨基酸组成统计、分子量、等电点、亲
3、疏水性、信号肽分析（跨膜区域） 、二级结构（含保守结构域）及三维结构分析，miR 分析，启动子分析等。其中抗原表位分析、多序列比对和进化分析等。三 The nature and function of myoglobin：Nature: myoglobin = a polypeptide chain + a prosthetic group polypeptide chain, composed of 153 amino acid residuesProsthetic group: heme prosthetic groupMolecular weight: 16, 700Function:
4、transport of oxygen and oxygen storage function in muscle tertiary structure of myoglobin四 Key word: 肌红蛋白（myoglobin）五 Implements and methods：采用 Bioedit和 DNAMAN分析软件工具和在线有关蛋白质和核酸方面的分析软件，分别进行核酸和蛋白质两个方面的分析。核酸分析首先从 pubmed核酸数据库中搜索到人的 Myoglobin核酸序列，分别进行序列的基本性质分析，限制性酶谱分析及其基因在染色体上的定位分析。具体操作方法将根据上述各个软件上“Help”
5、 文件进行，基因在染色体上的定位分析根据网页上的操作说明进行。从 pubmed蛋白质数据库中搜索到人的 Myoglobin和整个前体 Myoglobin蛋白质序列，分别进行蛋白序列的基本性质分析（氨基酸组成、分子量、等电点） ，亲疏水性分析，蛋白酶切位点分析，信号肽序列分析，二级结构和三维结构分析，同源序列和进化分析。二级结构分析包括结构功能域、Motif 、保守位点等，具体操作方法将根据上述各个软件上“Help ”文件和网页上的操作说明进行。Resluts:1.从 NCBI 中可以得到一下有关 Myoglobi蛋白分子的信息：Human myoglobin gene, exon 1GenBa
6、nk: M10090.1LOCUS HUMMGI1 2552 bp DNA linear PRI 07-JAN-1995DEFINITION Human myoglobin gene, exon 1.ACCESSION M10090VERSION M10090.1 GI:187580KEYWORDS Fok family repetitive sequence; direct repeat; myoglobin; repeatregion; tandem repeat.SEGMENT 1 of 3SOURCE Homo sapiens (human)ORGANISM Homo sapiensE
7、ukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini;Catarrhini; Hominidae; Homo.REFERENCE 1 (bases 1 to 2552)AUTHORS Akaboshi,E.TITLE Cloning of the human myoglobin geneJOURNAL Gene 33 (3), 241-249 (1985)PUBMED 2989088COMMENT Or
8、iginal source text: Human: fetal liver DNA (library of Lawn etal.), clone lambda-HMB151; delta-thalassemia leukocyte DNA (libraryof Kimura et al.), clone lambda-HMB159.Draft entry and computer-readable copy of sequence 1 kindlyprovided by E.Akaboshi, 22-OCT-1985.A tandem repeat sequence is found at
9、position 142-794. Position ofthe mRNA cap site was deduced from that of the seal myoglobin gene(Blanchetot et al. 1983).FEATURES Location/Qualifierssource 12552/organism=“Homo sapiens“/mol_type=“genomic DNA“/db_xref=“taxon:9606“/map=“22q11.2-qter“repeat_region 11001750/note=“Fok family repetitive el
10、ement“prim_transcript 18952552/note=“myoglobin mRNA and intron“gene join(19652552,M14602.1:12671,M14603.1:1501)/gene=“MB“CDS join(19652059,M14602.1:14151637,M14603.1:355501)/gene=“MB“/note=“myoglobin“/codon_start=1/protein_id=“AAA59595.1“/db_xref=“GI:386872“/translation=“MGLSDGEWQLVLNVWGKVEADIPGHGQE
11、VLIRLFKGHPETLEKFDKFKHLKSEDEMKASEDLKKHGATVLTALGGILKKKGHHEAEIKPLAQSHATKHKIPVKYLEFISECIIQVLQSKHPGDFGADAEGAMNKALELFRKDMASNYKELGFQG“exon 2552/gene=“MB“/note=“myoglobin intron A“ORIGIN 2 bp upstream of RsaI site.1 gtactgtatt ttcattcctc ttagttatct ccctaaaaag actctgagtt ccttgaacac61 aggaaggtgt tttatttgat tt
12、tgttatcc tcagcatgta gcagtgtctg acacacagta121 ggtgctctat cactgtgaga gggatggatg gatgggtgga gttacagatg gatagaagga181 tagatggagg gatgggtgga tgatggatgg atagatggat ggagggggga tgatgaatgg241 agggataatg agtggatgaa tgagggaatg ggtggatgga tggatggagg gatggaggaa301 cagatagata gatggaggga tgggtgggtg atggatggat agat
13、ggatgg agggagggat361 gatgaatgga gggataatga atggatgaat gaggggatgg gtggatggat gaatggaggg421 atgatgggtg gatgaatgaa ttgagggatg gatggatgaa cacatggatg gatggataga481 tggatagatg gaggaactgg tggattttgg atggatgggt ggatggatag atgaatgaat541 gcctggatag acaaagagat gatggataga tgaatagatg aattaaggga tgtcggatag601 atg
14、gagggat tgatagatgt tggatggatg ggtggtggat ggatagatga gtgaatgcat661 ggatagacaa agagatgatg gatggatgaa ttaagggatg acagatggat ggatggatga721 gtaactggat ggacaagtgg ataaatggat agatggttga atacctgaat ggattgaagg781 aggatgcatg gatgtaagat aaggctaatc atcctccact ctctttcttt gcaaaaccat841 ccacccattt actcaataaa cattt
15、attca gttcaaactt ggcacaaagc accatgtgag901 gcccaagaga tacgtgggtt aataaaacag agctcctgcc ctcctgaaaa ctgcaaagaa961 aggggcgtgg cttcctgagt tcaaatccca actctgccag cgactagctg tacatcagtg1021 atgtttccct actttctctc aattaaatag ggataatgtc agtacctatc acattgggag1081 gtcttgcggg gattaaatga gttaccaaat gccaagtgtt tggga
16、caggg cctggcaccc1141 agcaaagtct cttgtgagtg ctggctgcta ttatcctaat ggagaagatg gcatgaaaac1201 caggaaatag gatgcccttt gggaagcaat gcaacaggaa cttacacaaa gaaaggaaag1261 gaggaagcaa ttagtggtgt ctcaaaggag tatgtcaaga aaaacttttc agagggaaac1321 ctttgagcag ggtcatgaaa acaggagttc tctaagagat tgtggacttg cctgggacca1381
17、 cctggctata agcacaaaac catccggttc ctttctgtca cttctggcgg gtgaggggtc1441 tctggcaaag gggcagaagg tgcgtgagag gttgcgaatg gccaggactg tcctggggcc1501 agccggggca cctggtggcc aagcttagaa acatgacagg tcctcttggg agggctgacc1561 gcagggagcg ttgggtttca ggctgctggc gtcggcttct gtggtgccct ttctgtcggc1621 tatgagagtc cagacagt
18、gc ccaacctcct ccccttcttt ccacacgcac aaccacccca1681 ccccctgtgg cctgagctgt cctgcctcgc cacaatggca cctgccctaa aatagcttcc1741 catgtgaggg ctagagaaag gaaaagatta gaccctccct ggatgagaga gagaaagtga1801 aggagggcag gggaggggga cagcgagcca ttgagcgatc tttgtcaagc atcccagaag1861 gtataaaaac gcccttggga ccaggcagcc tcaaac
19、ccca gctgttgggg ccaggacacc1921 cagtgagccc atacttgctc tttttgtctt cttcagactg cgccatgggg ctcagcgacg1981 gggaatggca gttggtgctg aacgtctggg ggaaggtgga ggctgacatc ccaggccatg2041 ggcaggaagt cctcatcagg taaaaggaag agattccatt gcccctgcca cccacaccct2101 aagatcaagg gtgttcagct gcaaggtgga aagtttgcac gtggggtagg tcag
20、ttggct2161 gcattagtta agggtgttag aacggtcact tgctttttct ttgcttttaa gtgtcaggga2221 ttggactcag gagagggaaa ggagccattt caggctgatg tcagcagctg gaggaagcat2281 gagaatcaaa cctaggatgc tcagagtcca ccaggaagaa ttttagaatt atagacagtc2341 agagttaaca agggtcctga gagattttgt acagccacct ctcttacagg atgaggacaa2401 aaagcgact
21、g agaaggggag gacatttcca gagtcacagc tcattaaatg ctcttaaagt2461 gtcaaggtta agacatgctc ttcaagggga gacagatctg gttctagact tggctctgcc2521 actgagccac tgggtgacct ttgggaaggt acOfficial Symbol MBprovided by HGNCOfficial Full Name myoglobinprovided by HGNCPrimary source :HGNC:6915Locus tag :RP4-569D19.7Seerelat
22、ed：Ensembl:ENSG00000198125; HPRD:01170; MIM:160000; Vega:OTTHUMG00000150606Gene type:protein coding Refsep status:REVIEWEDOrganism： Homo sapiens Lineage： Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini; Catarrhini; Hominid
23、ae; HomoAlso known as：PVALBGenomic contextLocation:22q13.1Sequence:Chromosome: 22; NC_000022.10 (3600281136019401, complement)See MB in Epigenomics, MapViewerChromosome 22 - NC_000022.10Genomic regions, transcripts, and productsGenomicSequence NC_000022 chromosome 22 reference GRCh37.p13 Primary Ass
24、emblyGo to nucleotideGraphics FASTA GenBankNC_000022.10: 36M36M (22Kbp) C Find on Sequence: - + Tools Configure 2. 利用 Primer premier5.0 软件对 myoglobin 核酸序列进行引物设计其结果如下：Sense primer:5 TACTCGTGCCTTTCGTACTAGGCTC 3Anti-sense primer:5 ATGAGCACGGAAAGCATGATCCGAG 33. 蛋白质序列分析利用 ExPASy 软件包对该基因进行氨基酸组成统计、分子量、等电点等分析及磷酸化分析
