[原创]pPICHOLI 穿梭载体系统
<p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; TEXT-INDENT: 21pt; mso-char-indent-count: 2.0;"><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">pPICHOLI</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">载体可以让异源基因在</span><i><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">P. pastoris</font></span></i><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-style: italic;">中表达，同时也能在原核的</span><i><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">E.coli</font></span></i><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-style: italic;">中表达。这个载体包含可诱导（酵母）的乙醇脱氢酶（</span><span lang="EN-US" style="COLOR: black; mso-bidi-font-style: italic;"><font face="Times New Roman">AOX</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-style: italic;">）启动子和</span><span lang="EN-US" style="COLOR: black; mso-bidi-font-style: italic;"><font face="Times New Roman">E.coli T7</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-style: italic;">启动子，这样就允许它可以在</span><i><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">P. pastoris</font></span></i><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-style: italic;">和</span><i><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">E.coli</font></span></i><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-style: italic;">中自主复制。因此，载体的线性化就不再是必需的了，而且，少量的</span><span lang="EN-US" style="COLOR: black; mso-bidi-font-style: italic;"><font face="Times New Roman">DNA</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-style: italic;">就可成功的转化</span><i><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">P. pastoris</font></span></i><i><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">。</span><span lang="EN-US" style="COLOR: black;"><p></p></span></i></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; TEXT-INDENT: 21.75pt;"><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-size: 10.5pt;">由于</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><font face="Times New Roman">PARS</font></span><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-size: 10.5pt;">序列被整合到载体中，使从酵母中提取的质粒非常简单的实现复原。不需要将亚克隆转入一系列表达载体中，也不需对阳性菌株进行检测，而多克隆位点又使</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><font face="Times New Roman">DNA</font></span><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-size: 10.5pt;">片段很方便的连接到载体中。</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt;"><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-size: 10.5pt;">特点：</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;"><span lang="EN-US" style="mso-bidi-font-size: 10.5pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">1.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span><span style="FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman"; mso-bidi-font-size: 10.5pt;">原核表达系统是一个易于操作、花费少，并且可得到高水平异源蛋白产物；</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;"><span lang="EN-US" style="mso-bidi-font-size: 10.5pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">2.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span><i><span lang="EN-US" style="COLOR: black; FONT-FAMILY: Arial;">P. pastoris</span></i><span lang="EN-US" style="COLOR: black; FONT-FAMILY: Arial;">/pPICHOLI </span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: Arial; mso-hansi-font-family: Arial; mso-bidi-font-family: Arial;">系统是一个强有力的真核表达系统，由于结合了</span><span lang="EN-US" style="COLOR: black; FONT-FAMILY: Arial;">AOX</span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: Arial; mso-hansi-font-family: Arial; mso-bidi-font-family: Arial;">或</span><span lang="EN-US" style="COLOR: black; FONT-FAMILY: Arial;">CUP1</span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: Arial; mso-hansi-font-family: Arial; mso-bidi-font-family: Arial;">启动子，使其可以快速生长。</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;"><span lang="EN-US" style="mso-bidi-font-size: 10.5pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">3.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: Arial; mso-hansi-font-family: Arial; mso-bidi-font-family: Arial; mso-bidi-font-style: italic;">适合表达翻译后修饰的可溶性蛋白和不适宜在大肠杆菌中表达的蛋白（真核）（例如一些毒素蛋白）；</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;"><span lang="EN-US" style="mso-bidi-font-size: 10.5pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">4.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: Arial; mso-hansi-font-family: Arial; mso-bidi-font-family: Arial; mso-bidi-font-style: italic;">容易克隆和高的转化效率；</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt 18pt; TEXT-INDENT: -18pt; mso-list: l0 level1 lfo1; tab-stops: list 18.0pt;"><span lang="EN-US" style="mso-bidi-font-size: 10.5pt; mso-fareast-font-family: "Times New Roman";"><span style="mso-list: Ignore;"><font face="Times New Roman">5.<span style="FONT: 7pt "Times New Roman";"> </span></font></span></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: Arial; mso-hansi-font-family: Arial; mso-bidi-font-family: Arial; mso-bidi-font-style: italic;">方便的亲和纯化和重组蛋白的检测。</span><span lang="EN-US" style="mso-bidi-font-size: 10.5pt;"><p></p></span></p>
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