Difference between revisions of "Part:BBa K4623006"
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The mSA-linker is a modified version of the Basic linker, divided into two parts while retaining the mSA portion, and still plays a role in constructing an avidin-biotin affinity system. The C-terminus of the recombinant protein contains a segment with the N-terminal sequence of GP41-1 (BBa_K3308067), which can be connected to the N-terminal of GP41-1C of the Cut linker (BBa_K3308068). Additionally, a TrxA solubility-enhancing protein tag is added to the N-terminus to improve protein solubility for expression. The His-tag is used for purification and separation, while the thrombin site is employed to remove the tag after purification.
 
The mSA-linker is a modified version of the Basic linker, divided into two parts while retaining the mSA portion, and still plays a role in constructing an avidin-biotin affinity system. The C-terminus of the recombinant protein contains a segment with the N-terminal sequence of GP41-1 (BBa_K3308067), which can be connected to the N-terminal of GP41-1C of the Cut linker (BBa_K3308068). Additionally, a TrxA solubility-enhancing protein tag is added to the N-terminus to improve protein solubility for expression. The His-tag is used for purification and separation, while the thrombin site is employed to remove the tag after purification.
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https://static.igem.wiki/teams/4623/wiki/cs-part/cs-part/cs-part/cs1-3d.png
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==Cultivation, Purification and SDS-PAGE==
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===Induction Condition===
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<html>
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<head><!-- HTML头部开始标签 -->
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<style>
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    .image-container {
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      float: left; /* 图像容器向左浮动 */
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      width: 50%; /* 图像容器宽度占据页面的二分之一 */
 +
      text-align: center; /* 图像容器中的内容居中对齐 */
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    }
 +
 +
    .image-container img {
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      display: block; /* 图像显示为块级元素 */
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      margin: 0 auto; /* 上下居中，左右自动居中 */
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      border: 1px solid black;
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      max-width: 100%; /* 图像最大宽度为容器宽度 */
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    }
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 +
    .image-container figcaption {
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      text-align: center; /* 图注居中对齐 */
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    }
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 +
    .text-container {
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      float: left; /* 文字容器向左浮动 */
 +
      width: 50%; /* 文字容器宽度占据页面的二分之一 */
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    }
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 +
    /* 清除浮动 */
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    .clearfix::after {
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      content: "";
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      display: table;
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      clear: both;
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    }
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  </style>
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</head> <!-- HTML头部结束标签 -->
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<body> <!-- HTML主体开始标签 -->
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    <!-- 在这里添加网页内容，包括文本、图片、链接等 -->
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 +
</body> <!-- HTML主体结束标签 -->
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 +
</html> <!-- 后缀：HTML元素的结束标签 -->
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 08:11, 10 October 2023


Cut Silinker 1(mSA-linker), TrxA-His-thrombin-mSA-GP41-1

Cuttable Silinker (CS) is a new recombinant protein, in addition to connecting to the surface of silica, the core breakthrough is the ability to cope with a variety of microenvironments specific cut, release off and then play a role in the release of the drugs. CS is divided into three parts, which can be self-sheared through the intein, and finally connected together to form a complete recombinant protein. The three components include: mSA-intein peptide (CS1) for coupling to the target protein, SBP-intein peptide (CS3) for linking to silica, and changeable recognized cleavage site (CS2). In our proof-of-concept, we chose the PLGVR motif, which can be recognized by metallo-matrix proteases (MMPs) in tumor microenvironments and added intein to both ends to achieve interchangeable insertion connections. that enable interchangeable insertion junctions.

The mSA-linker is a modified version of the Basic linker, divided into two parts while retaining the mSA portion, and still plays a role in constructing an avidin-biotin affinity system. The C-terminus of the recombinant protein contains a segment with the N-terminal sequence of GP41-1 (BBa_K3308067), which can be connected to the N-terminal of GP41-1C of the Cut linker (BBa_K3308068). Additionally, a TrxA solubility-enhancing protein tag is added to the N-terminus to improve protein solubility for expression. The His-tag is used for purification and separation, while the thrombin site is employed to remove the tag after purification. cs1-3d.png

Cultivation, Purification and SDS-PAGE

Induction Condition

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 445
    Illegal AgeI site found at 505
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 909