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huojinlong9287銀蟲 (小有名氣)
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為了克隆版納微型豬近交系血管內(nèi)皮生長因子（VEGF）以及檢測該因子在各組織中的轉(zhuǎn)錄水平，提取了版納微型豬近交系18種組織的RNA，反轉(zhuǎn)錄為cDNA，利用RT-PCR方法擴增VEGF基因編碼區(qū)全長序列，進行克隆測序。利用序列翻譯的氨基酸與牛等8個物種的VEGF氨基酸序列構(gòu)建出物種系統(tǒng)進化樹，同時采用半定量RT-PCR方法分析VEGF基因在版納微型豬近交系18種組織中的轉(zhuǎn)錄水平。結(jié)果顯示，版納微型豬近交系VEGF基因編碼區(qū)全長573 bp（GenBank登錄號：JF831364），編碼190個氨基酸。氨基酸序列同源性分析表明，與牛等8個物種具有90%以上的相似性，分子系統(tǒng)進化表明與人的關(guān)系最近，其次為牛、羊和兔。多組織半定量RT-PCR研究表明VEGF mRNA在版納微型豬近交系的18個組織中均有轉(zhuǎn)錄水平的表達，其中在卵巢、脾臟、心臟、肺臟及皮膚中表達量較高，其他組織中表達量略低。 In order to isolate the vascular endothelial growth factor and detect its transcriptions in different tissues of Banna mini-pig inbred line, the RNAs of 18 kinds of tissues from Banna Mini-pig Inbred Line (BMI) were extracted, separately and reverse transcribed for cDNA. The gene coding region sequence of vascular endothelial growth factor was amplified through RT-PCR, then the purified product of PCR was sequenced by clone method. The phylogeny evolution tree was constructed among putative VEGF amino acid of BMI with those other eight species, such as cattle. Meanwhile, the semi-quantitative RT-PCR was used to analyse the transcription profile of VEGF gene in 18 kinds of tissues of BMI. The result showed the coding region of VEGF gene was 573 bp (GenBank accession number：JF831364), and coded 190 amino acids. The amino acid homology analysis showed it had over 90% homology with 8 species, such as human. Molecular phylogeny evolution indicated it had a closer relation with human, then with cattle, sheep and rabbit. Semi-quantitative RT-PCR analysis of mRNA from different tissues indicted that VEGF mRNA expressed in 18 tissues of BMI, and the expression level was higher in ovary, spleen, heart, lung and skin. |
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2樓2011-12-31 10:54:00
wyky2007136
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【答案】應(yīng)助回帖
huojinlong9287(金幣+50, 翻譯EPI+1): 2011-12-31 18:48:28
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僅供參考 In order to clone the vascular endothelial growth factor (VEGF) of Banna miniature pig inbred line and detect the transcription level of the factor in different tissues, the RNA of 18 kinds of tissues of the Banna miniature pig inbred line was extracted, then the extracted RNA was subjected to reverse transcription to form cDNA, and the gene encoding region entire sequence of the VEGF was amplified through a RT-PCR method, thereby carrying out cloning and sequencing. A species system cladogram was constructed by utilizing sequence-translated amino acids or the VEGF amino acid sequence of cattle and the like eight species, meanwhile, the transcription level of the VEGF gene in 18 kinds of tissues of the Banna miniature pig inbred line was analyzed by using a semi-quantitative RT-PCR method. The results showed that the entire length of the gene encoding region of the VEGF of the Banna miniature pig inbred line was 573 bp (GenBank accession number：JF831364), and 190 amino acids were encoded. The amino acid sequence homology analysis showed that it had homology more than 90% with the cattle and the like eight species, and the molecular system evolution indicated that it had a most close correlation with human being, followed by cattle, sheep and rabbit. The multi-tissue semi-quantitative RT-PCR analysis indicted that the mRNA of the VEGF was expressed in 18 tissues of the Banna miniature pig inbred line in a transcription level, and the expression level was higher in ovary, spleen, heart, lung and skin and a little lower in other tissues. |
3樓2011-12-31 14:18:21
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