[推荐]pPICHOLI 穿梭载体系统
<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";">穿梭表达载体</span><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";">和</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">pPICHOLI-HA</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">结合了真核和原核启动子元件。噬菌体</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">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";">启动子，含有主要衣壳蛋白的核糖体结合位点，促进细菌的表达，它被放在</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">P. pastoris</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">启动子的下游。强的乙醇脱氢酶启动子（</span><span lang="EN-US" style="COLOR: black;"><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";">）是被紧紧调控的，在含有葡萄糖的培养基中，蛋白表达是完全被抑制的，而生长在甲醇中，则可被最大限度的诱导。</span><span lang="EN-US" style="COLOR: black;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; TEXT-INDENT: 21.75pt;"><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">pPICHOLI-1</font></span><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;"><font face="Times New Roman">3579bp</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">）有两个</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">G bases</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">直接位于</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">Sal</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">位点的上游，</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">pPICHOLI-2</font></span><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;"><font face="Times New Roman">3578bp</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">）和</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">pPICHOLI-3</font></span><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;"><font face="Times New Roman">3577bp</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">）分别敲除一个和两个</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">G bases</font></span><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></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; TEXT-INDENT: 21.75pt;"><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">pPICHOLI-HA</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">载体含有一个</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">HA</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">（血球凝集素）抗原决定簇代替了生物素酰化序列。</span><span lang="EN-US" style="COLOR: black;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; TEXT-INDENT: 21.75pt;"><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">pPICHOLI-C</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">加进了</span><span style="FONT-FAMILY: 宋体; mso-bidi-font-size: 10.5pt; mso-hansi-font-family: "Times New Roman"; mso-font-kerning: 0pt; mso-bidi-font-family: 宋体; mso-ansi-language: ZH-CN;">酿酒酵母</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">CUP1<span style="mso-tab-count: 1;"> </span></font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">启动子（代替了</span><span lang="EN-US" style="COLOR: black;"><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";">启动子）。这个载体没有</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">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";">启动子，不适合原核蛋白的表达。</span><span lang="EN-US" style="COLOR: black;"><p></p></span></p><p class="MsoNormal" style="MARGIN: 0cm 0cm 0pt; TEXT-INDENT: 21.75pt;"><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">由于使用了共同的选择标记</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">zeocin</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">，这个穿梭载体仍然是小的（</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">~3.6</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">和</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">3.2kb</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">），因此，操作、克隆和转化都很方便。通过将</span><i style="mso-bidi-font-style: normal;"><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">Pichia</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";">特异的自主复制序列（</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">PARS1</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">）整合到</span><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";">载体中，线性化不再是必需的，转化效率增加到</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">10<sup>5</sup></font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">转化体</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">/ug 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";">。</span><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";">双重表达载体包括一个</span><span lang="EN-US" style="COLOR: black;"><font face="Times New Roman">RGS</font></span><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;"><font face="Times New Roman">His</font></span><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;"><font face="Times New Roman">6tag</font></span><span style="COLOR: black; FONT-FAMILY: 宋体; mso-ascii-font-family: "Times New Roman"; mso-hansi-font-family: "Times New Roman";">和一个生物素化序列，这样就使表达蛋白易于检测和快速的纯化。</span><span lang="EN-US" style="COLOR: black;"><p></p></span></p><table class="MsoNormalTable" cellspacing="0" cellpadding="0" width="520" border="0" style="BACKGROUND: whitesmoke; WIDTH: 390pt; BORDER-COLLAPSE: collapse;"><tbody><tr style="mso-yfti-irow: 0; mso-yfti-firstrow: yes;"><td valign="top" width="70" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 0.75pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 0.75pt; BACKGROUND: gainsboro; PADDING-BOTTOM: 0.75pt; BORDER-LEFT: #ece9d8; WIDTH: 52.5pt; PADDING-TOP: 0.75pt; BORDER-BOTTOM: #ece9d8;"><p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 14pt; TEXT-ALIGN: left; mso-pagination: widow-orphan;"><b><span lang="EN-US" style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial; mso-font-kerning: 0pt;">order#<p></p></span></b></p></td><td valign="top" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 0.75pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 0.75pt; BACKGROUND: gainsboro; PADDING-BOTTOM: 0.75pt; BORDER-LEFT: #ece9d8; PADDING-TOP: 0.75pt; BORDER-BOTTOM: #ece9d8;"><p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 14pt; TEXT-ALIGN: left; mso-pagination: widow-orphan;"><b><span lang="EN-US" style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial; mso-font-kerning: 0pt;">description<p></p></span></b></p></td><td valign="top" width="100" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 0.75pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 0.75pt; BACKGROUND: gainsboro; PADDING-BOTTOM: 0.75pt; BORDER-LEFT: #ece9d8; WIDTH: 75pt; PADDING-TOP: 0.75pt; BORDER-BOTTOM: #ece9d8;"><p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt; LINE-HEIGHT: 14pt; TEXT-ALIGN: left; mso-pagination: widow-orphan;"><b><span lang="EN-US" style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial; mso-font-kerning: 0pt;">amount*<p></p></span></b></p></td></tr><tr style="mso-yfti-irow: 1; mso-yfti-lastrow: yes;"><td valign="top" width="70" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 0.75pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 0.75pt; PADDING-BOTTOM: 0.75pt; BORDER-LEFT: #ece9d8; WIDTH: 52.5pt; PADDING-TOP: 0.75pt; BORDER-BOTTOM: gainsboro 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid gainsboro .75pt;"><p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt; TEXT-ALIGN: left; mso-pagination: widow-orphan;"><span lang="EN-US" style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial; mso-font-kerning: 0pt;">PPICH<p></p></span></p></td><td valign="top" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 0.75pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 0.75pt; PADDING-BOTTOM: 0.75pt; BORDER-LEFT: #ece9d8; PADDING-TOP: 0.75pt; BORDER-BOTTOM: gainsboro 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid gainsboro .75pt;"><p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt; TEXT-ALIGN: left; mso-pagination: widow-orphan;"><span lang="EN-US" style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial; mso-font-kerning: 0pt;">pPICHOLI-1 vector DNA<br/>pPICHOLI-2 vector DNA<br/>pPICHOLI-3 vector DNA<br/>pPICHOLI-C vector DNA<br/>pPICHOLI-HA vector DNA<br/>sequencing primers<p></p></span></p></td><td valign="top" width="100" style="BORDER-RIGHT: #ece9d8; PADDING-RIGHT: 0.75pt; BORDER-TOP: #ece9d8; PADDING-LEFT: 0.75pt; PADDING-BOTTOM: 0.75pt; BORDER-LEFT: #ece9d8; WIDTH: 75pt; PADDING-TOP: 0.75pt; BORDER-BOTTOM: gainsboro 1pt solid; BACKGROUND-COLOR: transparent; mso-border-bottom-alt: solid gainsboro .75pt;"><p class="MsoNormal" align="left" style="MARGIN: 0cm 0cm 0pt; TEXT-ALIGN: left; mso-pagination: widow-orphan;"><span lang="EN-US" style="FONT-SIZE: 10pt; COLOR: black; FONT-FAMILY: Arial; mso-font-kerning: 0pt;">10 µg<br/>10 µg<br/>10 µg<br/>10 µg<br/>10 µg<br/>500 pmole each<p></p></span></p></td></tr></tbody></table>
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