本文選題：纖連蛋白(FN) + 錨蛋白重復(fù)(ANK)　； 參考：《四川農(nóng)業(yè)大學(xué)學(xué)報(bào)》2017年03期

【摘要】：【目的】獲得豬FANK1基因CDS序列、組織表達(dá)和蛋白質(zhì)功能信息。【方法】以GenBank下載的豬及近緣物種的FANK1 mRNA序列為參考序列,設(shè)計(jì)特異引物從版納微型豬近交系(BMI)睪丸組織中擴(kuò)增FANK1基因完全編碼序列及部分側(cè)翼序列。應(yīng)用實(shí)時(shí)熒光定量PCR技術(shù)檢測(cè)BMI 15個(gè)重要組織的FANK1 mRNA轉(zhuǎn)錄表達(dá)水平,并對(duì)FANK1翻譯的蛋白質(zhì)序列進(jìn)行多種功能生物信息學(xué)分析,預(yù)測(cè)FANK1蛋白質(zhì)的功能,最后構(gòu)建FANK1多物種氨基酸系統(tǒng)進(jìn)化樹(shù)�！窘Y(jié)果】獲得了BMI FANK1編碼區(qū)序列長(zhǎng)1 041 bp,編碼346個(gè)氨基酸,序列已提交GenBank,基因登錄號(hào)為KU705617和KU705618,對(duì)應(yīng)的氨基酸登錄號(hào)為AOC89035和AOC89036�；蚪M結(jié)構(gòu)分析表明FANK1基因定位于豬14號(hào)染色體,有11個(gè)外顯子和10個(gè)內(nèi)含子。多組織相對(duì)熒光定量表達(dá)分析表明FANK1基因在睪丸、尿道球腺和精囊腺中呈高表達(dá)水平;在其他組織中呈中低表達(dá)水平。功能生物信息學(xué)分析表明FANK1蛋白質(zhì)含有2種保守結(jié)構(gòu)域FN3和ANK,無(wú)跨膜螺旋結(jié)構(gòu),無(wú)N端信號(hào)肽序列,二級(jí)結(jié)構(gòu)以無(wú)規(guī)卷曲為主,N末端和C末端均親水,有4類(lèi)功能活性位點(diǎn)。系統(tǒng)進(jìn)化分析表明,FANK1基因在進(jìn)化中高度保守,且與牛、羊的親緣關(guān)系最近,符合物種的系統(tǒng)分類(lèi)學(xué)�！窘Y(jié)論】成功克隆了版納微型豬近交系FANK1基因的CDS序列并進(jìn)行了多種組織表達(dá)分析,為后續(xù)研究FANK1基因在小型豬精細(xì)胞分裂及調(diào)節(jié)信號(hào)通路方面的作用及功能奠定基礎(chǔ)。
[Abstract]:[objective] to obtain the CDS sequence, tissue expression and protein function information of porcine FANK1 gene. [methods] the FANK1 mRNA sequence of porcine and related species downloaded by GenBank was used as reference sequence.A specific primer was designed to amplify the complete coding sequence and partial flanking sequence of FANK1 gene from BMI testis of Banna miniature pig inbred line.The transcription and expression of FANK1 mRNA in 15 important tissues of BMI were detected by real-time fluorescence quantitative PCR, and the protein sequences translated by FANK1 were analyzed by multifunctional bioinformatics to predict the function of FANK1 protein.Finally, the phylogenetic tree of FANK1 multi-species amino acids was constructed. [results] the coding region of BMI FANK1 was 1 041 BP, encoding 346 amino acids. The sequence was submitted to GenBank, the gene accession numbers were KU705617 and KU705618, and the corresponding amino acid login numbers were AOC89035 and AOC89036.Genomic structure analysis showed that FANK1 gene was located on pig chromosome 14 with 11 exons and 10 introns.Quantitative analysis of relative fluorescence in multiple tissues showed that the expression of FANK1 gene was high in testis urethra glandular gland and seminal vesicle gland and low expression level in other tissues.Functional bioinformatics analysis showed that the FANK1 protein contained two conserved domains, FN3 and ANK, no transmembrane helical structure, no N-terminal signal peptide sequence, and the secondary structure was mainly composed of random curls, which were hydrophilic at the N-terminal and C-terminal, and had four functional active sites.Phylogenetic analysis showed that the FANK1 gene was highly conserved in evolution and had a close relationship with cattle and sheep.[conclusion] the CDS sequence of the FANK1 gene of Banna miniature pig inbred line was cloned successfully and the expression of various tissues was analyzed.The results provide a basis for further study on the role and function of FANK1 gene in spermatocyte division and regulation of signaling pathway in miniature pigs.
【作者單位】： 華中農(nóng)業(yè)大學(xué)生命科學(xué)技術(shù)學(xué)院;云南農(nóng)業(yè)大學(xué)動(dòng)物科學(xué)技術(shù)學(xué)院;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31660637;31460580;31660650)
【分類(lèi)號(hào)】：Q78;S828

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