家蠶絲氨酸蛋白酶基因BmSP25轉(zhuǎn)錄分析及其免疫響應(yīng)
發(fā)布時(shí)間：2018-11-19 13:57
【摘要】：【目的】分析家蠶絲氨酸蛋白酶(SP)基因序列BmSP25及轉(zhuǎn)錄情況,明確其表達(dá)規(guī)律對防御家蠶核型多角體病毒(BmNPV)入侵的免疫應(yīng)答機(jī)制,為揭示BmSPs在家蠶免疫應(yīng)答方面的功能作用提供理論依據(jù)�！痉椒ā靠寺mSP25基因序列,對該基因編碼蛋白的氨基酸序列、分子量、結(jié)構(gòu)域等進(jìn)行生物信息學(xué)分析;利用Gene Doc和MEGA5.0對BmSP25氨基酸序列進(jìn)行多序列比對及系統(tǒng)發(fā)育進(jìn)化樹分析,采用半定量RT-PCR對家蠶不同組織和發(fā)育時(shí)期的BmSP25基因轉(zhuǎn)錄情況進(jìn)行分析,并以實(shí)時(shí)熒光定量PCR檢測BmSP25基因在BmNPV感染家蠶中腸組織中的轉(zhuǎn)錄水平�！窘Y(jié)果】BmSP25基因的ORF全長885 bp,編碼294個(gè)氨基酸,其中第1~17位氨基酸為信號肽,去信號肽的分子量為29.1 kD,理論等電點(diǎn)為7.8。BmSP25蛋白由4個(gè)α螺旋、15個(gè)β折疊和一些無規(guī)則卷曲構(gòu)成;其氨基酸序列同源性比對分析發(fā)現(xiàn),BmSP25(BGIBMGA008514-PA)與蓓帶夜蛾SP序列(Gen Bank登錄號ADM35105)的同源性最高,為62.1%。BmSP25基因在家蠶中腸組織中特異表達(dá),且在整個(gè)幼蟲時(shí)期呈持續(xù)性表達(dá)。BmSP25基因在家蠶感染BmNPV后發(fā)生明顯變化,至感染6 h時(shí)呈下調(diào)趨勢,而在感染3、12和24 h時(shí)均呈明顯上調(diào)表達(dá)�！窘Y(jié)論】BmSP25在防御BmNPV入侵家蠶的免疫應(yīng)答過程中發(fā)揮重要作用。鑒于昆蟲SP具有高度保守的底物特異性位點(diǎn),因此可利用底物類似物、基因定點(diǎn)突變等方式來預(yù)防農(nóng)林害蟲。
[Abstract]:[objective] to analyze the sequence and transcription of serine protease (SP) gene in Bombyx mori (Bombyx mori), and to clarify the immune response mechanism of its expression to (BmNPV) invasion of Bombyx mori nuclear polyhedrosis virus. [methods] the sequence of BmSP25 gene was cloned, and the amino acid sequence, molecular weight and domain of the protein were analyzed by bioinformatics. Gene Doc and MEGA5.0 were used to analyze the amino acid sequences of BmSP25 and phylogenetic tree, and semi-quantitative RT-PCR was used to analyze the transcription of BmSP25 gene in different tissues and developmental stages of Bombyx mori (Bombyx mori). The transcriptional level of BmSP25 gene in the midgut of silkworm infected with BmNPV was detected by real-time fluorescence quantitative PCR. [results] the total length of ORF of BmSP25 gene encoded 294 amino acids, in which the first 17 amino acid was a signal peptide. The molecular weight of the de-signalling peptide is 29.1 kD,. The theoretical isoelectric point of 7.8.BmSP25 protein is composed of four 偽 helix, 15 尾 folds and some irregular curls. The homology analysis of amino acid sequence showed that BmSP25 (BGIBMGA008514-PA) had the highest homology with SP (Gen Bank accession number ADM35105), which indicated that 62.1%.BmSP25 gene was specifically expressed in the midgut of silkworm, Bombyx mori (Bombyx mori). The expression of BmSP25 gene in silkworm was significantly changed after infection of BmNPV, and decreased at 6 h after infection. The expression of BmSP25 was upregulated significantly at 12 and 24 h after infection. [conclusion] BmSP25 plays an important role in the immune response to BmNPV invasion of Bombyx mori (Bombyx mori). In view of the highly conserved substrate-specific sites of insect SP, substrates analogue and gene site-directed mutation can be used to prevent agricultural and forestry pests.
【作者單位】： 云南省農(nóng)業(yè)科學(xué)院蠶桑蜜蜂研究所;
【基金】：國家蠶桑產(chǎn)業(yè)技術(shù)體系建設(shè)專項(xiàng)項(xiàng)目(CARS-22) 云南省現(xiàn)代農(nóng)業(yè)蠶桑產(chǎn)業(yè)技術(shù)體系建設(shè)項(xiàng)目(2013KJTX006) 云南農(nóng)業(yè)科學(xué)院蠶桑蜜蜂研究所青年創(chuàng)新基金項(xiàng)目(QC2015004)
【分類號】：S884.51
[Abstract]:[objective] to analyze the sequence and transcription of serine protease (SP) gene in Bombyx mori (Bombyx mori), and to clarify the immune response mechanism of its expression to (BmNPV) invasion of Bombyx mori nuclear polyhedrosis virus. [methods] the sequence of BmSP25 gene was cloned, and the amino acid sequence, molecular weight and domain of the protein were analyzed by bioinformatics. Gene Doc and MEGA5.0 were used to analyze the amino acid sequences of BmSP25 and phylogenetic tree, and semi-quantitative RT-PCR was used to analyze the transcription of BmSP25 gene in different tissues and developmental stages of Bombyx mori (Bombyx mori). The transcriptional level of BmSP25 gene in the midgut of silkworm infected with BmNPV was detected by real-time fluorescence quantitative PCR. [results] the total length of ORF of BmSP25 gene encoded 294 amino acids, in which the first 17 amino acid was a signal peptide. The molecular weight of the de-signalling peptide is 29.1 kD,. The theoretical isoelectric point of 7.8.BmSP25 protein is composed of four 偽 helix, 15 尾 folds and some irregular curls. The homology analysis of amino acid sequence showed that BmSP25 (BGIBMGA008514-PA) had the highest homology with SP (Gen Bank accession number ADM35105), which indicated that 62.1%.BmSP25 gene was specifically expressed in the midgut of silkworm, Bombyx mori (Bombyx mori). The expression of BmSP25 gene in silkworm was significantly changed after infection of BmNPV, and decreased at 6 h after infection. The expression of BmSP25 was upregulated significantly at 12 and 24 h after infection. [conclusion] BmSP25 plays an important role in the immune response to BmNPV invasion of Bombyx mori (Bombyx mori). In view of the highly conserved substrate-specific sites of insect SP, substrates analogue and gene site-directed mutation can be used to prevent agricultural and forestry pests.
【作者單位】： 云南省農(nóng)業(yè)科學(xué)院蠶桑蜜蜂研究所;
【基金】：國家蠶桑產(chǎn)業(yè)技術(shù)體系建設(shè)專項(xiàng)項(xiàng)目(CARS-22) 云南省現(xiàn)代農(nóng)業(yè)蠶桑產(chǎn)業(yè)技術(shù)體系建設(shè)項(xiàng)目(2013KJTX006) 云南農(nóng)業(yè)科學(xué)院蠶桑蜜蜂研究所青年創(chuàng)新基金項(xiàng)目(QC2015004)
【分類號】：S884.51
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