基因工程作业（引物设计） - 图文

口腔鳞癌组织中肿瘤转移相关基因1（MTA1）

一、选择原因及应用

口腔鳞癌组织中肿瘤转移相关基因1（MTA1）在蛋白和mRNA的表达水平，揭示其与口腔鳞癌（OSCC）发生、发展的关系。方法采用免疫组织化学法和原位杂交技术检测46例OSCC标本、15例口腔黏膜白斑与20例正常口腔黏膜标本中MTAl基因的表达水平，并分析其与OSCC临床病理学参数的关系。结果MTA1蛋白和MTA1mRNA在OSCC组织中的表达水平显著高于口腔黏膜白斑和正常口腔黏膜（P〈0．05），口腔黏膜白斑中MTA1蛋白和MTA1mRNA表达水平显著高于口腔正常黏膜（P〈0．01），MTA1蛋白和MTA1mRNA表达与肿瘤浸润深度和淋巴结转移密切相关（P〈0．05）。结论MTA1基因在蛋白和mRNA的表达水平在OSCC发生、发展及浸润转移过程中起一定促进作用，有望成为判断OSCC预后及选择治疗方案的一个新肿瘤标志物。

二、2查阅NCBI得到MTA1相关信息并获得目的基因

PREDICTED: Gorilla gorilla gorilla metastasis associated 1 (MTA1), mRNA

NCBI Reference Sequence: XM_004055801.1

FASTA Graphics

LOCUS XM_004055801 2872 bp mRNA linear PRI 03-DEC-2012

DEFINITION PREDICTED: Gorilla gorilla gorilla metastasis associated 1 (MTA1), mRNA.

ACCESSION XM_004055801

VERSION XM_004055801.1 GI:426378238 KEYWORDS .

SOURCE Gorilla gorilla gorilla (western lowland gorilla) ORGANISM Gorilla gorilla gorilla Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi;

Mammalia; Eutheria; Euarchontoglires; Primates; Haplorrhini;

Catarrhini; Hominidae; Gorilla.

COMMENT MODEL REFSEQ: This record is predicted by automated computational

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analysis. This record is derived from a genomic sequence

(NW_004005914) annotated using gene prediction method: GNOMON,

supported by mRNA and EST evidence. Also see:

Documentation of NCBI's Annotation Process

##Genome-Annotation-Data-START## Annotation Provider :: NCBI

Annotation Status :: Full annotation

Annotation Version :: Gorilla gorilla Annotation Release 100

Annotation Pipeline :: NCBI eukaryotic genome annotation pipeline

Annotation Method :: Best-placed RefSeq; Gnomon Features Annotated :: Gene; mRNA; CDS; ncRNA ##Genome-Annotation-Data-END## FEATURES Location/Qualifiers source 1..2872

/organism=\ /mol_type=\

/sub_species=\ /db_xref=\9595\ /chromosome=\ gene 1..2872

/gene=\

/note=\analysis using

gene prediction method: GNOMON. Supporting evidence

includes similarity to: 4 mRNAs, 303 ESTs, 7 Proteins\ /db_xref=\101134898\根据 misc_feature 104..106 /gene=\ /note=\ CDS 212..2359 /gene=\MTA1\ /codon_start=1

/product=\MTA1\

/protein_id=\XP_004055849.1\ /db_xref=\

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/db_xref=\101134898\

/translation=\

FYRRRDISSTLIALADKHATLSVCYKAGPGADNGEEGEIEEEMENPEMVDLPEKLKHQ

LRHRELFLSRQLESLPATHIRGKCSVTLLNETESLKSYLEREDFFFYSLVYDPQQKTL

LADKGEIRVGNRYQADITDLLKEGEEDGRDQSKLETKVWEAHNPLTDKQIDQFLVVAR

SVGTFARALDCSSSVRQPSLHMSAAAASRDITLFHAMDTLHKNIYDISKAISALVPQG

GPVLCRDEMEEWSASEANLFEEALEKYGKDFTDIQQDFLPWKSLTSIIEYYYMWKTTD

RYVQQKRLKAAEAESKLKQVYIPNYNKPNPNQISVNNVKAGVVNGTGAPGQSPGAGRA

CESCYTTQSYQWYSWGPPNMQCRLCASCWTYWKKYGGLKMPTRLDGERPGPNRSNMSP

HGLPARSSGSPKFAMKTRQAFYLHTTKLTRIARRLCREILRPWHAARHPYLPINSAAI

KAECTARLPEASQSPLVLKQAVRKPLEAVLRYLETHPRPPKPDPVKSVSSVLSSLTPA

KVAPVINNGSPTILGKRSYEQHNGVDGNMKKRLLMPSRGLANHGQTRHMGPSRNLLLN

GKSYPTKVRLIRGGSLPPVKRRRMNWIDAPDDVFYMATEETRKIRKLLSSSETKRAAR RPYKPIALRQSQALPLRPPPPAPVNDEPIVIED\ORIGIN

1 tcctcctctt cctctcccgc ccgcgccgcg gccctcccgt ccctgcgcgg cctcggcggc

61 ctcggcggcg gcggcggcgg cggcggcagc agcgcggccc ctttaaacgc ctgcggcgcc

121 ccccgccccc gccatcgcgc ctccattttc ccggccgccc gcgccgagcg ccgcgcccgc 起始密码子 181 cccgggcccc tccgccgccg ccggcccgga catggccgcc aacatgtaca gggtcggaga

241 ctacgtctac tttgagaact cctccagcaa cccatacctg atccggagga tcgaggagct

301 caacaagacg gccaatggga acgtggaggc caaagtggtg tgcttctacc ggaggcggga

361 catctccagc accctcatcg ccctggccga caagcacgca accctgtcag tctgctataa

421 ggccggaccg ggggcggaca acggcgagga aggggaaata gaagaggaaa tggagaatcc

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481 ggaaatggtg gacctgcccg agaaactaaa gcaccagctg cggcatcggg agctgttcct

541 ctcccggcag ctggagtctc tgcccgccac gcacatcagg ggcaagtgca gcgtcaccct

601 gctcaacgag accgagtcgc tcaagtccta cctggagcgg gaggatttct tcttctattc

661 tctagtctac gacccacagc agaagaccct gctggcagat aaaggagaga ttcgagtagg

721 aaaccggtac caggcagaca tcaccgactt gttaaaagaa ggcgaggagg atggccgaga

781 ccagtccaag ttggagacca aggtgtggga ggcgcacaac ccactcacag acaagcagat

841 cgaccagttc ctggtggtgg cccgctctgt gggcaccttc gcacgggccc tggactgcag

901 cagctccgtc cgacagccca gcctgcacat gagcgccgca gctgcctccc gagacatcac

961 gctgttccac gccatggata ctctccacaa gaacatctat gacatctcca aggccatctc

1021 ggcactggtg ccgcagggcg ggcccgtgct ctgcagggac gagatggagg agtggtctgc

1081 atcagaggcc aaccttttcg aggaagccct ggaaaaatat gggaaggatt tcacggacat

1141 tcagcaagat tttctcccgt ggaagtcgct gaccagcatc attgagtact actacatgtg

1201 gaagaccacc gacagatacg tgcagcagaa acgcttgaaa gcagctgaag ctgagagcaa

1261 gttaaagcaa gtttatattc ccaactataa caagccaaat ccgaaccaaa tcagtgtcaa

1321 caacgtcaag gccggtgtgg tgaatggcac gggggcgccg ggccagagcc ctggggctgg

1381 ccgggcctgc gagagctgtt acaccacaca gtcttaccag tggtattctt ggggtccccc

1441 taacatgcag tgtcgtctct gcgcatcttg ttggacatat tggaagaaat atggtggctt

1501 gaaaatgcca acccggttag atggagagag gccaggacca aaccgcagta acatgagtcc

1561 ccacggcctc ccagcccgga gcagcgggag ccccaagttt gccatgaaga ccaggcaggc

1621 tttctatctg cacacgacga agctgacgcg gatcgcccgg cgcctgtgcc gtgagatcct

1681 gcgcccgtgg cacgctgcgc ggcaccccta cctgcccatc aacagtgcgg ccatcaaggc

1741 cgagtgcacg gcgcggctgc ccgaagcctc ccagagcccg ctggtgctga agcaggcggt

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1801 acgcaagccg ctggaagccg tgcttcggta tcttgagacc cacccccgtc cccccaagcc

1861 tgaccccgtg aaaagcgtgt ccagcgtgct cagcagcctg acgcccgcca aggtggcccc

1921 cgtcatcaac aacggctccc ccaccatcct gggcaagcgc agctacgagc agcacaacgg

1981 ggtggacggc aacatgaaga agcgcctctt gatgcccagt aggggtctgg caaaccacgg

2041 acagaccagg cacatgggac caagccggaa cctcctgctc aacgggaagt cctaccccac

2101 caaagtgcgc ctgatccggg ggggctccct gcccccagtc aagcggcggc ggatgaactg

2161 gatcgacgcc ccggatgacg tgttctacat ggccacagag gagaccagga agatccgcaa

2221 gctgctctca tcctcggaaa ccaagcgtgc tgcccgccgg ccctacaagc ccatcgccct

2281 gcgccagagc caggccctgc cgctgcggcc accgccacct gcgcccgtca acgacgagcc 终止密码子 2341 catcgtcatc gaggactagg ggccgccccc acctgcggcc gccccccgcc cctcgcccgc

2401 ccacacggcc ccttcccagc cagcccgccg cccgcccctc agtttggtag tgccccacct

2461 cccgccctca cctgcagaga aacgcgctcc ttggcggaca ctgagggagg agaggaagaa

2521 gcgcggctaa cttattccga gaatgccgag gagttgtcgt ttttagcttt gtgtttactt

2581 tttggctgga gcggagatga ggggccaccc cgtgcccctg tgctgcgggg ccttttgccc

2641 ggaggccggg ccctaaggtt ttgttgtgtt ctgttgaagg tgccgtttta aattttattt

2701 ttattacttt ttttgtagat gaacttgagc tctgtaactt acacctggaa tgttaggatc

2761 gtgcggccgc ggccggccga gctgcctggc ggggttggcc cttgtctttt caagtaattt

2821 tcatattaaa caaaaacaaa gaaaaaaatc ttataaaaag gaaaaaaacc aa //

三、表达载体的选择

我所选用的原核表达载体为质粒pBR322；

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