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huojinlong9287银虫 (小有名气)
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为了克隆版纳微型猪近交系血管内皮生长因子（VEGF）以及检测该因子在各组织中的转录水平，提取了版纳微型猪近交系18种组织的RNA，反转录为cDNA，利用RT-PCR方法扩增VEGF基因编码区全长序列，进行克隆测序。利用序列翻译的氨基酸与牛等8个物种的VEGF氨基酸序列构建出物种系统进化树，同时采用半定量RT-PCR方法分析VEGF基因在版纳微型猪近交系18种组织中的转录水平。结果显示，版纳微型猪近交系VEGF基因编码区全长573 bp（GenBank登录号：JF831364），编码190个氨基酸。氨基酸序列同源性分析表明，与牛等8个物种具有90%以上的相似性，分子系统进化表明与人的关系最近，其次为牛、羊和兔。多组织半定量RT-PCR研究表明VEGF mRNA在版纳微型猪近交系的18个组织中均有转录水平的表达，其中在卵巢、脾脏、心脏、肺脏及皮肤中表达量较高，其他组织中表达量略低。 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楼2011-12-31 10:54:00
wyky2007136
金虫 (小有名气)
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huojinlong9287(金币+50, 翻译EPI+1): 2011-12-31 18:48:28
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仅供参考 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楼2011-12-31 14:18:21