1、1广州大学学生实验报告开课学院及实验室：生命科学学院 计机楼 407 2013 年 11 月 15 日 学 院 生命科学学院 年级、专业、班 生物工程 10 姓名 学号 10142000实验课程名称 生物信息学 成绩 实验项目名称 实验三 序列分析与比较 指 导 教 师 一、实验目的1、掌握已知或未知序列接受号的核酸序列检索的基本步骤；2、掌握使用 BioEdit 软件进行核酸序列的基本分析；3 熟悉基于核酸序列比对分析的真核基因结构分析（内含子/外显子分析）；4、了解基因的电子表达谱分析。二、实验内容1、归纳对人瘦素 (leptin) 的核酸序列分析的结果，列出主要的分析结果；2、总结核酸序
2、列分析的基本步骤，相互对比结果，指出应注意的事项。2三、实验步骤3.1、人瘦素 (leptin) 的核酸序列分析Homo sapiens leptin (LEP), mRNANCBI Reference Sequence: NM_000230.2FASTA GraphicsGo to:LOCUS NM_000230 3444 bp mRNA linear PRI 03-NOV-2013DEFINITION Homo sapiens leptin (LEP), mRNA.ACCESSION NM_000230VERSION NM_000230.2 GI:169790920KEYWORDS Ref
3、Seq.SOURCE Homo sapiens (human)ORGANISM Homo sapiensEukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini;Catarrhini; Hominidae; Homo.REFERENCE 1 (bases 1 to 3444)AUTHORS Lee,J.H., Zhou,L., Kwon,K.S., Lee,D., Park,B.H. and Kim,J.
4、R.TITLE Role of leptin in Legg-Calve-Perthes diseaseJOURNAL J. Orthop. Res. 31 (10), 1605-1610 (2013)PUBMED 23832827REMARK GeneRIF: Leptin might play an important role in Legg-Calve-Perthes3disease pathogenesis.REFERENCE 2 (bases 1 to 3444)AUTHORS Fourati,M., Mnif,M., Kharrat,N., Charfi,N., Kammoun,
5、M., Fendri,N.,Sessi,S., Abid,M., Rebai,A. and Fakhfakh,F.TITLE Association between Leptin gene polymorphisms and plasma leptinlevel in three consanguineous families with obesityJOURNAL Gene 527 (1), 75-81 (2013)PUBMED 23751306REMARK GeneRIF: Two functional variants in the regulatory region of theLEP
6、 gene are associated with plasma leptin level as a quantitativetrait.REFERENCE 3 (bases 1 to 3444)AUTHORS Miranda-Filloy,J.A., Lopez-Mejias,R., Genre,F., Carnero-Lopez,B.,Ochoa,R., Diaz de Teran,T., Gonzalez-Juanatey,C., Blanco,R.,Llorca,J. and Gonzalez-Gay,M.A.TITLE Leptin and visfatin serum levels
7、 in non-diabetic ankylosingspondylitis patients undergoing TNF-alpha antagonist therapyJOURNAL Clin. Exp. Rheumatol. 31 (4), 538-545 (2013)PUBMED 23711190REMARK GeneRIF: In non-diabetic patients with ankylosing spondylitis ontreatment with infliximab leptin and visfatin serum levels do notcorrelate
8、with disease activity or systemic inflammation.REFERENCE 4 (bases 1 to 3444)AUTHORS Gulcelik,N.E., Halil,M., Ariogul,S. and Usman,A.TITLE Adipocytokines and aging: adiponectin and leptinJOURNAL Minerva Endocrinol. 38 (2), 203-210 (2013)4PUBMED 23732375REMARK GeneRIF: we briefly discuss the role of t
9、wo major adipocytokinesadiponectin and leptin in the aging process and age-relateddiseases-REVIEWReview articleREFERENCE 5 (bases 1 to 3444)AUTHORS Lakhdar,N., Ben Saad,H., Denguezli,M., Zaouali,M., Zbidi,A.,Tabka,Z. and Bouassida,A.TITLE Effects of intense cycling training on plasma leptin andadipo
10、nectin and its relation to insulin resistanceJOURNAL Neuro Endocrinol. Lett. 34 (3), 229-235 (2013)PUBMED 23685422REMARK GeneRIF: Adiponectin is sensitive to maximal exercise when realisedafter intense training and that six months heavy cycling trainingdont affect adiponectin concentrations, but dec
11、reases thesynthesis of leptin and homostasis and improves aerobic capacity.REFERENCE 6 (bases 1 to 3444)AUTHORS Isse,N., Ogawa,Y., Tamura,N., Masuzaki,H., Mori,K., Okazaki,T.,Satoh,N., Shigemoto,M., Yoshimasa,Y., Nishi,S. et al.TITLE Structural organization and chromosomal assignment of the humanobe
12、se geneJOURNAL J. Biol. Chem. 270 (46), 27728-27733 (1995)PUBMED 7499240REFERENCE 7 (bases 1 to 3444)AUTHORS Masuzaki,H., Ogawa,Y., Isse,N., Satoh,N., Okazaki,T., Shigemoto,M.,Mori,K., Tamura,N., Hosoda,K., Yoshimasa,Y. et al.5TITLE Human obese gene expression. Adipocyte-specific expression andregio
13、nal differences in the adipose tissueJOURNAL Diabetes 44 (7), 855-858 (1995)PUBMED 7789654REFERENCE 8 (bases 1 to 3444)AUTHORS Zhang,Y., Proenca,R., Maffei,M., Barone,M., Leopold,L. andFriedman,J.M.TITLE Positional cloning of the mouse obese gene and its human homologueJOURNAL Nature 372 (6505), 425
14、-432 (1994)PUBMED 7984236REMARK Erratum:Nature 1995 Mar 30;374(6521):479REFERENCE 9 (bases 1 to 3444)AUTHORS Friedman,J.M., Leibel,R.L., Siegel,D.S., Walsh,J. and Bahary,N.TITLE Molecular mapping of the mouse ob mutationJOURNAL Genomics 11 (4), 1054-1062 (1991)PUBMED 1686014REFERENCE 10 (bases 1 to
15、3444)AUTHORS Groot,P.H., Scholte,H.R. and Hulsmann,W.C.TITLE Fatty acid activation: specificity, localization, and functionJOURNAL Adv. Lipid Res. 14, 75-126 (1976)PUBMED 3952COMMENT REVIEWED REFSEQ: This record has been curated by NCBI staff. Thereference sequence was derived from DA762132.1, U4365
16、3.1,AC018635.10 and BU752306.1.This sequence is a reference standard in the RefSeqGene project.On Mar 12, 2008 this sequence version replaced gi:4557714.6Summary: This gene encodes a protein that is secreted by whiteadipocytes, and which plays a major role in the regulation of bodyweight. This prote
17、in, which acts through the leptin receptor,functions as part of a signaling pathway that can inhibit foodintake and/or regulate energy expenditure to maintain constancy ofthe adipose mass. This protein also has several endocrinefunctions, and is involved in the regulation of immune andinflammatory r
18、esponses, hematopoiesis, angiogenesis and woundhealing. Mutations in this gene and/or its regulatory regions causesevere obesity, and morbid obesity with hypogonadism. This gene hasalso been linked to type 2 diabetes mellitus development. providedby RefSeq, Jul 2008.Sequence Note: This RefSeq record
19、 was created from transcript andgenomic sequence data to make the sequence consistent with thereference genome assembly. The genomic coordinates used for thetranscript record were based on transcript alignments.Publication Note: This RefSeq record includes a subset of thepublications that are availa
20、ble for this gene. Please see the Generecord to access additional publications.#Evidence-Data-START#Transcript exon combination : U43653.1, BC060830.1 ECO:0000332RNAseq introns : single sample supports all introns7ERS025081, ERS025084 ECO:0000348#Evidence-Data-END#COMPLETENESS: full length.PRIMARY R
21、EFSEQ_SPAN PRIMARY_IDENTIFIER PRIMARY_SPAN COMP1-309 DA762132.1 1-309310-1807 U43653.1 309-18061808-3383 AC018635.10 34398-359733384-3444 BU752306.1 1-61 cFEATURES Location/Qualifierssource 13444/organism=“Homo sapiens“/mol_type=“mRNA“/db_xref=“taxon:9606“/chromosome=“7“/map=“7q31.3“gene 13444/gene=
22、“LEP“/gene_synonym=“LEPD; OB; OBS“/note=“leptin“/db_xref=“GeneID:3952“/db_xref=“HGNC:6553“/db_xref=“HPRD:01249“/db_xref=“MIM:164160“exon 129/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“8/inference=“alignment:Splign:1.39.8“STS 7611/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/db_xref=“UniSTS:481242“exon 30201/
23、gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/inference=“alignment:Splign:1.39.8“CDS 58561/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/note=“leptin (murine obesity homolog); leptin (obesityhomolog, mouse); obese, mouse, homolog of; obesity factor;obese protein“/codon_start=1/product=“leptin precursor“/protein
24、_id=“NP_000221.1“/db_xref=“GI:4557715“/db_xref=“CCDS:CCDS5800.1“/db_xref=“GeneID:3952“/db_xref=“HGNC:6553“/db_xref=“HPRD:01249“/db_xref=“MIM:164160“/translation=“MHWGTLCGFLWLWPYLFYVQAVPIQKVQDDTKTLIKTIVTRINDISHTQSVSSKQKVTGLDFIPGLHPILTLSKMDQTLAVYQQILTSMPSRNVIQISNDLE9NLRDLLHVLAFSKSCHLPWASGLETLDSLGGVLEA
25、SGYSTEVVALSRLQGSLQDMLWQLDLSPGC“sig_peptide 58120/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/inference=“COORDINATES: ab initio prediction:SignalP:4.0“mat_peptide 121558/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/product=“leptin“STS 172254/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/standard_name=“LEP-3“/db_xre
26、f=“UniSTS:253879“exon 2023427/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/inference=“alignment:Splign:1.39.8“STS 594843/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/standard_name=“STS-U43653“/db_xref=“UniSTS:35281“STS 594699/gene=“LEP“10/gene_synonym=“LEPD; OB; OBS“/standard_name=“OBS“/db_xref=“UniSTS:35280“
27、STS 703898/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/standard_name=“RH70631“/db_xref=“UniSTS:78152“STS 31933281/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“/standard_name=“D8S2279“/db_xref=“UniSTS:473907“polyA_signal 34053410/gene=“LEP“/gene_synonym=“LEPD; OB; OBS“polyA_site 3427/gene=“LEP“/gene_synonym=“L
28、EPD; OB; OBS“ORIGIN 1 gtaggaatcg cagcgccagc ggttgcaagg cccaagaagc ccatcctggg aaggaaaatg61 cattggggaa ccctgtgcgg attcttgtgg ctttggccct atcttttcta tgtccaagct121 gtgcccatcc aaaaagtcca agatgacacc aaaaccctca tcaagacaat tgtcaccagg181 atcaatgaca tttcacacac gcagtcagtc tcctccaaac agaaagtcac cggtttggac241 ttc
29、attcctg ggctccaccc catcctgacc ttatccaaga tggaccagac actggcagtc11301 taccaacaga tcctcaccag tatgccttcc agaaacgtga tccaaatatc caacgacctg361 gagaacctcc gggatcttct tcacgtgctg gccttctcta agagctgcca cttgccctgg421 gccagtggcc tggagacctt ggacagcctg gggggtgtcc tggaagcttc aggctactcc481 acagaggtgg tggccctgag cag
30、gctgcag gggtctctgc aggacatgct gtggcagctg541 gacctcagcc ctgggtgctg aggccttgaa ggtcactctt cctgcaagga ctacgttaag601 ggaaggaact ctggcttcca ggtatctcca ggattgaaga gcattgcatg gacacccctt661 atccaggact ctgtcaattt ccctgactcc tctaagccac tcttccaaag gcataagacc721 ctaagcctcc ttttgcttga aaccaaagat atatacacag gatcc
31、tattc tcaccaggaa781 gggggtccac ccagcaaaga gtgggctgca tctgggattc ccaccaaggt cttcagccat841 caacaagagt tgtcttgtcc cctcttgacc catctccccc tcactgaatg cctcaatgtg901 accaggggtg atttcagaga gggcagaggg gtaggcagag cctttggatg accagaacaa961 ggttccctct gagaattcca aggagttcca tgaagaccac atccacacac gcaggaactc1021 cca
32、gcaacac aagctggaag cacatgttta tttattctgc attttattct ggatggattt1081 gaagcaaagc accagcttct ccaggctctt tggggtcagc cagggccagg ggtctccctg1141 gagtgcagtt tccaatccca tagatgggtc tggctgagct gaacccattt tgagtgactc1201 gagggttggg ttcatctgag caagagctgg caaaggtggc tctccagtta gttctctcgt1261 aactggtttc atttctactg t
33、gactgatgt tacatcacag tgtttgcaat ggtgttgccc1321 tgagtggatc tccaaggacc aggttatttt aaaaagattt gttttgtcaa gtgtcatatg1381 taggtgtctg cacccagggg tggggaatgt ttgggcagaa gggagaagga tctagaatgt1441 gttttctgaa taacatttgt gtggtgggtt ctttggaagg agtgagatca ttttcttatc1501 ttctgcaatt gcttaggatg tttttcatga aaatagctct
34、 ttcagggggg ttgtgaggcc1561 tggccaggca ccccctggag agaagtttct ggccctggct gaccccaaag agcctggaga1621 agctgatgct ttgcttcaaa tccatccaga ataaaacgca aagggctgaa agccatttgt1681 tggggcagtg gtaagctctg gctttctccg actgctaggg agtggtcttt cctatcatgg1741 agtgacggtc ccacactggt gactgcgatc ttcagagcag gggtccttgg tgtgaccc
35、tc1801 tgaatggtcc agggttgatc acactctggg tttattacat ggcagtgttc ctatttgggg121861 cttgcatgcc aaattgtagt tcttgtctga ttggctcacc caagcaaggc caaaattacc1921 aaaaatcttg gggggttttt actccagtgg tgaagaaaac tcctttagca ggtggtcctg1981 agacctgaca agcactgcta ggcgagtgcc aggactcccc aggccaggcc accaggatgg2041 cccttcccac
36、tggaggtcac attcaggaag atgaaagagg aggtttgggg tctgccacca2101 tcctgctgct gtgtttttgc tatcacacag tgggtggtgg atctgtccaa ggaaacttga2161 atcaaagcag ttaactttaa gactgagcac ctgcttcatg ctcagccctg actggtgcta2221 taggctggag aagctcaccc aataaacatt aagattgagg cctgccctca gggatcttgc2281 attcccagtg gtcaaaccgc actcaccca
37、t gtgccaaggt ggggtattta ccacagcagc2341 tgaacagcca aatgcatggt gcagttgaca gcaggtggga aatggtatga gctgaggggg2401 gccgtgccca ggggcccaca gggaaccctg cttgcacttt gtaacatgtt tacttttcag2461 ggcatcttag cttctattat agccacatcc ctttgaaaca agataactga gaatttaaaa2521 ataagaaaat acataagacc ataacagcca acaggtggca ggaccag
38、gac tatagcccag2581 gtcctctgat acccagagca ttacgtgagc caggtaatga gggactggaa ccagggagac2641 cgagcgcttt ctggaaaaga ggagtttcga ggtagagttt gaaggaggtg agggatgtga2701 attgcctgca gagagaagcc tgttttgttg gaaggtttgg tgtgtggaga tgcagaggta2761 aaagtgtgag cagtgagtta cagcgagagg cagagaaaga agagacagga gggcaagggc2821 c
39、atgctgaag ggaccttgaa gggtaaagaa gtttgatatt aaaggagtta agagtagcaa2881 gttctagaga agaggctggt gctgtggcca gggtgagagc tgctctggaa aatgtgaccc2941 agatcctcac aaccacctaa tcaggctgag gtgtcttaag ccttttgctc acaaaacctg3001 gcacaatggc taattcccag agtgtgaaac ttcctaagta taaatggttg tctgtttttg3061 taacttaaaa aaaaaaaaaa
40、 aagtttggcc gggtgcggtg gctcacgcct gtaatcccag3121 cactttggga ggccaaggtg gggggatcac aaggtcacta gatggcgagc atcctggcca3181 acatggtgaa accccgtctc tactaaaaac acaaaagtta gctgagcgtg gtggcgggcg3241 cctgtagtcc cagccactcg ggaggctgag acaggagaat cgcttaaacc tgggaggcgg3301 agagtacagt gagccaagat cgcgccactg cactccgg
41、cc tgatgacaga gcgagattcc3361 gtcttaaaaa aaaaaaaaaa aaagtttgtt tttaaaaaaa tctaaataaa ataactttgc133421 cccctgcaaa aaaaaaaaaa aaaa/gi|169790920|ref|NM_000230.2| Homo sapiens leptin (LEP), mRNAGTAGGAATCGCAGCGCCAGCGGTTGCAAGGCCCAAGAAGCCCATCCTGGGAAGGAAAATGCATTGGGGAACCCTGTGCGGATTCTTGTGGCTTTGGCCCTATCTTTTCTAT
42、GTCCAAGCTGTGCCCATCCAAAAAGTCCAAGATGACACCAAAACCCTCATCAAGACAATTGTCACCAGGATCAATGACATTTCACACACGCAGTCAGTCTCCTCCAAACAGAAAGTCACCGGTTTGGACTTCATTCCTGGGCTCCACCCCATCCTGACCTTATCCAAGATGGACCAGACACTGGCAGTCTACCAACAGATCCTCACCAGTATGCCTTCCAGAAACGTGATCCAAATATCCAACGACCTGGAGAACCTCCGGGATCTTCTTCACGTGCTGGCCTTCTCTAAGAGCTGCCAC
43、TTGCCCTGGGCCAGTGGCCTGGAGACCTTGGACAGCCTGGGGGGTGTCCTGGAAGCTTCAGGCTACTCCACAGAGGTGGTGGCCCTGAGCAGGCTGCAGGGGTCTCTGCAGGACATGCTGTGGCAGCTGGACCTCAGCCCTGGGTGCTGAGGCCTTGAAGGTCACTCTTCCTGCAAGGACTACGTTAAGGGAAGGAACTCTGGCTTCCAGGTATCTCCAGGATTGAAGAGCATTGCATGGACACCCCTTATCCAGGACTCTGTCAATTTCCCTGACTCCTCTAAGCCACTCTTCCAAAGG
44、CATAAGACCCTAAGCCTCCTTTTGCTTGAAACCAAAGATATATACACAGGATCCTATTCTCACCAGGAAGGGGGTCCACCCAGCAAAGAGTGGGCTGCATCTGGGATTCCCACCAAGGTCTTCAGCCATCAACAAGAGTTGTCTTGTCCCCTCTTGACCCATCTCCCCCTCACTGAATGCCTCAATGTGACCAGGGGTGATTTCAGAGAGGGCAGAGGGGTAGGCAGAGCCTTTGGATGACCAGAACAAGGTTCCCTCTGAGAATTCCAAGGAGTTCCATGAAGACCACATCCACACACG
45、CAGGAACTCCCAGCAACACAAGCTGGAAGCACATGTTTATTTATTCTGCATTTTATTCTGGATGGATTTGAAGCAAAGCACCAGCTTCTCCAGGCTCTTTGGGGTCAGCCAGGGCCAGGGGTCTCCCTGGAGTGCAGTTTCCAATCCCATAGATGGGTCTGGCTGAGCTGAACCCATTTTGAGTGACTCGAGGGTTGGGTTCATCTGAGCAAGAGCTGGCAAAGGTGGCTCTCCAGTTAGTTCTCTCGTAACTGGTTTCATTTCTACTGTGACTGATGTTACATCACAGTGTTTGCAATG
46、GTGTTGCCCTGAGTGGATCTCCAAGGACCAGGTTATTTTAAAAAGATTTGTTTTGTCAAGTGTCATATGTAGGTGTCTGCACCCAGGGGTGGGGAATGTTTGGGCAGAAGGGAGAAGGATCTAGAATGTGTTTTCTGAATAACATTTGTGTGGTGGGTTCTTTGGAAGGAGTGAGATCATTTTCTTATCTTCTGCAATTGCTTAGGATGTTTTTCATGAAAATAGCTCT14TTCAGGGGGGTTGTGAGGCCTGGCCAGGCACCCCCTGGAGAGAAGTTTCTGGCCCTGGCTGACCCCAAA
47、GAGCCTGGAGAAGCTGATGCTTTGCTTCAAATCCATCCAGAATAAAACGCAAAGGGCTGAAAGCCATTTGTTGGGGCAGTGGTAAGCTCTGGCTTTCTCCGACTGCTAGGGAGTGGTCTTTCCTATCATGGAGTGACGGTCCCACACTGGTGACTGCGATCTTCAGAGCAGGGGTCCTTGGTGTGACCCTCTGAATGGTCCAGGGTTGATCACACTCTGGGTTTATTACATGGCAGTGTTCCTATTTGGGGCTTGCATGCCAAATTGTAGTTCTTGTCTGATTGGCTCACCCAAGCAAGG
48、CCAAAATTACCAAAAATCTTGGGGGGTTTTTACTCCAGTGGTGAAGAAAACTCCTTTAGCAGGTGGTCCTGAGACCTGACAAGCACTGCTAGGCGAGTGCCAGGACTCCCCAGGCCAGGCCACCAGGATGGCCCTTCCCACTGGAGGTCACATTCAGGAAGATGAAAGAGGAGGTTTGGGGTCTGCCACCATCCTGCTGCTGTGTTTTTGCTATCACACAGTGGGTGGTGGATCTGTCCAAGGAAACTTGAATCAAAGCAGTTAACTTTAAGACTGAGCACCTGCTTCATGCTCAGCCCTGACTGGTGCTATAGGCTGGAGAAGCTCACCCAATAAACATTAAGATTGAGGCCTGCCCTCAGGGATCTTGCATTCCCAGTGGTCAAACCGCACTCACCCATGTGCCAAGGTGGGGTATTTACCACAGCAGCTGAACAGCCAAATGCATGGTGCAGTTGACAGCAGGTGGGAAATGGTATGAGCTGAGGGGGGCCGTGCCCAGGGGCCCACAGGGAACCCTGCTTGCACTTTGTAACATGTTTACTTTTCAGGGCATCTTAGCTTCTATTATAGCCACATCCCTTTGAAACAAGA
