松墨天牛ADP核糖基化因子1基因的序列及表達特性分析
發(fā)布時間:2018-04-16 09:01
本文選題:松墨天牛 + ADP核糖基化因子 ; 參考:《河南農(nóng)業(yè)科學》2017年07期
【摘要】:為了探討Ras基因超家族中ADP核糖基化因子1基因(ARF1)在昆蟲生長發(fā)育進程中以及在溫度脅迫下的表達特性,以松墨天牛為研究對象,從已構建的cDNA文庫中篩選到松墨天牛ARF1基因,命名為MaARF1(Gen Bank:KY368168),對其進行序列分析以及不同蟲態(tài)和不同溫度脅迫下的表達特性分析。結果顯示,MaARF1基因編碼區(qū)序列長為549 bp,編碼182個氨基酸。預測蛋白質二級結構主要由α螺旋與β片層組成,其次是無規(guī)則卷曲和β轉角。通過DNAMAN軟件比對發(fā)現(xiàn),MaARF1與赤擬谷盜ARF1蛋白氨基酸序列完全一致,二者同源性為100%,且存在1個十四;稽c(G~2)和5個保守域(G_1 box—G_5 box)。通過RT-qPCR分析表明,MaARF1的表達量與松墨天牛的完全變態(tài)發(fā)育相關,各蟲態(tài)不間斷表達,幼蟲期在2齡幼蟲(L2)、5齡幼蟲(L5)中表達量較高,蛹化期間(L5—P0)表達量表現(xiàn)為上升趨勢,并在第2天蛹(P2)中達到最大值,羽化期間(P10—A0)表達量表現(xiàn)為下降趨勢;MaARF1在不同溫度脅迫下均有表達,低溫與高溫都會導致表達量下調(diào),溫度越高或越低,表達量下調(diào)越顯著。因此,MaARF1可能主要影響松墨天牛蛹化和羽化這2個變態(tài)發(fā)育關鍵點,從而影響整個變態(tài)發(fā)育歷程,同時其也可能參與了松墨天牛體內(nèi)依賴溫度的Ca~(2+)信號途徑。
[Abstract]:In order to investigate the expression of ADP - ribosylation factor - 1 gene ( ARF1 ) in the development of insect and its expression in the course of growth and development of insect , it was screened from the constructed cDNA library . The results showed that the sequence length of MaARF1 gene was 549 bp and 182 amino acids were encoded . The results showed that the sequence length of MaARF1 gene was 549 bp and 182 amino acids were encoded . The predicted protein secondary structure was composed mainly of 偽 - helix and 尾 - slice , and the homology was 100 % , and there were 1 myristoylation site ( G ~ 2 ) and 5 conserved domains ( G _ 1 box 鈥,
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