發(fā)布時間：2018-01-26 01:09

  本文關(guān)鍵詞： 沙梨 休眠 DELLA蛋白 PpGAI 原核表達　出處：《果樹學報》2017年02期 　論文類型：期刊論文

【摘要】：【目的】分離沙梨DELLA蛋白編碼基因PpGAI3,探究其蛋白在花芽休眠中的表達情況�！痉椒ā繌牟煌枥淞可忱嫫贩N‘蜜雪梨’和‘黃花梨’中分離PpGAI3基因,重組到p EASY-Blunt E1表達載體中,轉(zhuǎn)化BL21(DE3)宿主菌后經(jīng)IPTG誘導表達,并對表達菌株進行SDS-PAGE和Western blotting檢測�！窘Y(jié)果】獲得2種沙梨的PpGAI3基因,分別命名為PpGAI3mx和PpGAI3hh,Gen Bank登錄號為KU954535和KX078215,其開放閱讀框(ORF)為1 641 bp,編碼546個氨基酸,預測其為親水性不穩(wěn)定蛋白。進化樹分析表明PpGAI3mx和PpGAI3hh與白梨的親緣關(guān)系最近;成功構(gòu)建了p EASY-Blunt E1-PpGAI3原核表達載體,得到蛋白分子質(zhì)量約60 ku的誘導表達產(chǎn)物�！窘Y(jié)論】所得沙梨DELLA蛋白編碼基因PpGAI3為植物DELLA蛋白家族中的成員,在梨花芽休眠過程中具有特異表達,且表達產(chǎn)物大部分以不溶性包涵體形式存在。
[Abstract]:[objective] to isolate the DELLA protein encoding gene PpGAI3. To investigate the expression of its protein in flower bud dormancy. [methods] PpGAI3 gene was isolated from the varieties' Honeyue 'and' Huanghua pear'. The expression was induced by IPTG after recombinant into the expression vector of p EASY-Blunt E1 and transformed into BL21DDE3. SDS-PAGE and Western blotting were used to detect the expression strain. [results] the PpGAI3 gene of two kinds of pear was obtained. The login numbers for PpGAI3mx and PpGAI3hhlgen Bank are KU954535 and KX078215, respectively. Its open reading frame (ORF) was 1 641 BP, encoding 546 amino acids. Phylogenetic tree analysis showed that PpGAI3mx and PpGAI3hh had the closest phylogenetic relationship with white pear. The prokaryotic expression vector of p EASY-Blunt E1-PpGAI3 was successfully constructed. The protein molecular weight was about 60 ku. [conclusion] the DELLA protein encoding gene PpGAI3 is a member of the plant DELLA protein family. There was a specific expression in pear bud dormancy, and most of the expressed products existed in the form of insoluble inclusion body.
【作者單位】： 福建農(nóng)林大學園藝學院·福建農(nóng)林大學園藝植物天然產(chǎn)物研究所;
【基金】：國家自然科學基金(31501694) 高等學校博士學科點專項科研基金(20113515110011)
【分類號】：S661.2
【正文快照】： 沙梨(Pyrus pyrifolia)為福建省主要落葉果樹,具有冬季芽休眠的有益特性,喜溫暖濕潤環(huán)境,適應能力強,休眠期短,在我國南方地區(qū)被大面積推廣[1]。近年來在引進‘翠冠’‘翠玉’等新品種進行更新?lián)Q代的同時,仍缺少短低溫梨優(yōu)良品種,制約了福建亞熱帶氣候?qū)嫔a(chǎn)優(yōu)勢的發(fā)揮[2-3]，

本文編號：1464221