CN112513276B - 蛋白质异二聚体及其用途 - Google Patents

蛋白质异二聚体及其用途 Download PDF

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CN112513276B
CN112513276B CN201980050580.6A CN201980050580A CN112513276B CN 112513276 B CN112513276 B CN 112513276B CN 201980050580 A CN201980050580 A CN 201980050580A CN 112513276 B CN112513276 B CN 112513276B
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张晋宇
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Abstract

蛋白质异二聚体及其用途，蛋白质异二聚体包含第一多肽链以及不同于第一多肽链的第二多肽链，其中第一多肽链包含IL12a以及与IL12a融合的第一因子，第二多肽链包含IL12b以及与IL12b融合的第二因子，且其中第一因子及第二因子各自独立地选自下组：IL2、GMCSF、IL7、IL15、IL21和FLT3L。蛋白质异二聚体可用于治疗肿瘤。

Description

蛋白质异二聚体及其用途
技术领域
本申请涉及肿瘤治疗领域，具体而言，本申请涉及蛋白质异二聚体及其用途。
背景技术
肿瘤是一种严重威胁人类健康的疾病，近年来，免疫治疗作为一种新疗法，在肿瘤治疗中显示出了巨大的潜力。细胞因子(Cytokine)是体内非常重要的免疫信号，细胞因子融合蛋白技术是当今肿瘤免疫疗法的另一个热点方向。该方法是基于这些细胞因子具有相同或相关的功能活性而各自作用靶点不同，利用基因工程技术将两种或多种细胞因子融合在一起。但是目前利用细胞因子融合蛋白技术进行肿瘤治疗的效果仍不尽人意，有较多需要改进之处。
发明内容
本申请提供了一种蛋白质异二聚体及其用途，所述蛋白质异二聚体至少具有抗肿瘤效果好的有益效果。
一方面，本申请提供了一种蛋白质异二聚体，其包含第一多肽链以及不同于所述第一多肽链的第二多肽链，其中所述第一多肽链包含IL12a以及与所述IL12a融合的第一因子，所述第二多肽链包含IL12b以及与所述IL12b融合的第二因子，且其中所述第一因子及所述第二因子各自独立地选自下组：IL2、GMCSF、IL7、IL15、IL21和FLT3L。
在某些实施方式中，所述第一因子与所述第二因子不同。
在某些实施方式中，所述第一因子和所述第二因子彼此不同地分别选自细胞因子组A和细胞因子组B，所述细胞因子组A选自下组：IL2、IL7、IL15和IL21，且细胞因子组B选自下组：GMCSF和FLT3L。
在某些实施方式中，在所述第一多肽链中，所述IL12a的C末端与所述第一因子的N末端直接或间接融合。在某些实施方式中，在所述第一多肽链中，所述IL12a的N末端与所述第一因子的C末端直接或间接融合。在某些实施方式中，在所述第二多肽链中，所述IL12b的C末端与所述第二因子的N末端直接或间接融合。在某些实施方式中，在所述第二多肽链中，所述IL12b的N末端与所述第二因子的C末端直接或间接融合。
在某些实施方式中，所述第一因子和所述第二因子选自下组：
1)第一因子为IL2，且第二因子为GMCSF；
2)第一因子为IL7，且第二因子为GMCSF；
3)第一因子为IL15，且第二因子为GMCSF；
4)第一因子为IL21，且第二因子为GMCSF；
5)第一因子为IL2，且第二因子为FLT3L；
6)第一因子为IL7，且第二因子为FLT3L；
7)第一因子为IL15，且第二因子为FLT3L；
8)第一因子为IL21，且第二因子为FLT3L；
9)第一因子为GMCSF，且第二因子为IL2；
10)第一因子为GMCSF，且第二因子为IL7；
11)第一因子为GMCSF，且第二因子为IL15；
12)第一因子为GMCSF，且第二因子为IL21；
13)第一因子为FLT3L，且第二因子为IL2；
14)第一因子为FLT3L，且第二因子为IL7；
15)第一因子为FLT3L，且第二因子为IL15；以及，
16)第一因子为FLT3L，且第二因子为IL21。
在某些实施方式中，所述IL12a、所述IL12b、所述第一因子和所述第二因子来源自哺乳动物。
在某些实施方式中，所述IL12a、所述IL12b、所述第一因子和所述第二因子的来源相同。
在某些实施方式中，所述第一多肽链包括如SEQ ID NO.60-65和SEQ ID NO.70-73中任一项所示的序列。
在某些实施方式中，所述第二多肽链包括如SEQ ID NO.66-69和SEQ ID NO.74-75中任一项所示的序列。
在某些实旌方式中，所述蛋白质异二聚体包括如SEQ ID NO.18-37中任一项所示的序列。
另一方面，本申请提供一种或多种分离的核酸分子，其编码所述的蛋白质异二聚体。
在某些实施方式中，所述核酸分子包括如SEQ ID NO.38-57中任一项所示的序列。
另一方面，本申请提供一种表达载体，其包括编码所述的蛋白质异二聚体的核苷酸序列。
另一方面，本申请提供一种宿主细胞，所述宿主细胞包括所述的表达载体。
另一方面，本申请提供一种药物组合物，所述药物组合物包括所述的蛋白质异二聚体，所述的核酸分子、所述的载体和/或所述的宿主细胞，以及药学上可接受的载体。
另一方面，本申请提供一种制备所述的蛋白质异二聚体的方法，其包括以下的步骤：在能够表达所述的蛋白质异二聚体的情况下，培养所述的宿主细胞。
另一方面，本申请提供一种所述的蛋白质异二聚体和所述的药物组合物在制备治疗肿瘤药物中的应用。
在某些实施方式中，所述肿瘤包括黑色素瘤。
本领域技术人员能够从下文的详细描述中容易地洞察到本申请的其它方面和优势。下文的详细描述中仅显示和描述了本申请的示例性实施方式。如本领域技术人员将认识到的，本申请的内容使得本领域技术人员能够对所公开的具体实施方式进行改动而不脱离本申请所涉及发明的精神和范围。相应地，本申请的附图和说明书中的描述仅仅是示例性的，而非为限制性的。
附图说明
本申请所涉及的发明的具体特征如所附权利要求书所显示。通过参考下文中详细描述的示例性实施方式和附图能够更好地理解本申请所涉及发明的特点和优势。对附图简要说明书如下：
图1显示所述肿瘤的生长情况；
图2显示所述蛋白质异二聚体的结构示意图；
图3显示所述蛋白质异二聚体的表达情况；
图4显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图5显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图6显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图7显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图8显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图9显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图10显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图11显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图12显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图13显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图14显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图15显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图16显示所述蛋白质异二聚体诱导肿瘤消退的情况；
图17显示所述蛋白质异二聚体的构建情况。
具体实施方式
以下由特定的具体实施例说明本申请发明的实施方式，熟悉此技术的人士可由本说明书所公开的内容容易地了解本申请发明的其他优点及效果。
在本申请中，术语“融合蛋白”通常指由两种或更多种多肽构成的蛋白质。在所述融合蛋白中，所述多肽可以具备不同的特性，该特性可以是生物学特性，如体外或体内活性；也可以是简单的化学特性或物理特性，如与靶分子结合、催化反应等。在所述融合蛋白中，所述多肽可以直接结合，也可以通过接头(例如，肽连接子)或间隔物(spacer)而间接结合所述融合蛋白可以由融合基因表达，在融合基因中编码一种多肽的核苷酸序列可以在框内附有编码与该多肽不同的另一多肽的核苷酸序列。所述融合基因可以由重组宿主细胞表达为单一的蛋白质。
在本申请中，术语“细胞因子融合蛋白”通常是指包含两种以上细胞因子的融合蛋白。所述细胞因子融合蛋白可以通过基因重组技术获得。所述细胞因子融合蛋白可以既具有所包含的细胞因子的独特的生物学活性，或者使其中某些活性显著提高，还可以通过生物学活性的互补及协同效应发挥出较单一细胞因子简单配伍所不具备的复合生物学功能，甚至还可能会产生一些新的结构及生物学功能。在本申请中，所述细胞因子融合蛋白可以为蛋白质异二聚体。
在本申请中，术语“蛋白质异二聚体”是由两个不同的多肽链组成的二聚体。在本申请中，所述蛋白质异二聚体可以包括第一多肽链和第二多肽链，所述第一多肽链不同于所述第二多肽链，其中所述第一多肽链可包含IL12a以及与所述IL12a融合的第一因子，所述第二多肽链可包含IL12b以及与所述IL12b融合的第二因子。
在本申请中，术语“细胞因子”通常是指主要由免疫细胞分泌的、能调节细胞功能的小分子蛋白质。所述细胞因子对于细胞间相互作用、细胞的生长和分化有重要调节作用。例如，所述细胞因子可以选自下组：白细胞介素和集落刺激因子。在本申请中，所述白细胞介素是由多种细胞产生并作用于多种细胞的一类细胞因子。白细胞介素在传递信息，激活与调节免疫细胞，介导T、B细胞活化、增殖与分化及在炎症反应中发挥重要的作用。例如，所述白细胞介素可以选自下组中的一种或多种：IL12、IL2、IL15、IL21和IL7。在本申请中，所述集落刺激因子可以为可刺激不同的造血干细胞在半固体培养基中形成细胞集落的细胞因子。所述集落刺激因子对不同发育阶段的造血干细胞起促增殖、分化的作用。例如，根据集落刺激因子的作用范围，可以将所述集落刺激因子分别命名为粒细胞集落刺激因子(G-CSF)、巨噬细胞集落刺激因子(M-CSF)、粒细胞和巨噬细胞集落刺激因子(GM-CSF)和多能集落刺激因子(multi-CSF，IL3)。例如，所述集落刺激因子可以选自下组中的一种或多种：FMS相关酪氨酸激酶3配体(FTL3L)和粒细胞巨噬细胞集落刺激因子(GMCSF)。
在本申请中，术语“IL12”通常是指白细胞介素-12。在本申请中，IL12和IL-12可以互换使用。IL12可刺激活化型T细胞增殖，促进Th0细胞细胞向Th1细胞分化；也可诱导CTL和NK细胞的细胞毒活性并促进其分泌IFN-γ、TNF-α、GM-CSF等细胞因子；或者促进NK细胞和IL-2Rα受体及CD56分子的表达，增强对肿瘤细胞的ADCC效应。IL12为异二聚体，其包括p40亚基(40kd)和p35亚基(35kd)，这两个亚基可以通过二硫键相连接。例如，小鼠IL12(mIL12)中的p35亚基可包含SEQ ID NO：1所示的氨基酸序列，p40亚基可包含SEQ ID NO：2所示的氨基酸序列。又例如，人IL12(hIL12)中的p35亚基可包含SEQ ID NO：9所示的氨基酸序列，p40亚基可包含SEQ ID NO：10所示的氨基酸序列。
在本申请中，术语“IL2”通常是指白细胞介素-2。IL2对机体的免疫应答和抗病毒感染等有重要作用，能刺激已被特异性抗原启动的T细胞增殖；同时IL2能活化T细胞，促进细胞因子产生；刺激NK细胞增殖，增强NK杀伤活性及产生细胞因子，诱导LAK细胞产生；并且促进B细胞增殖和分泌抗体；激活巨噬细胞。例如，小鼠IL2(mIL2)可包含SEQ ID NO：3所示的氨基酸序列。又例如，人细胞因子IL2(hIL2)可包含SEQ ID NO：11所示的氨基酸序列。
在本申请中，术语“IL15”通常是指白细胞介素-15。IL15由活化的单核-巨噬细胞、表皮细胞和成纤维细胞等多种细胞产生，其可诱导B细胞增殖和分化。例如，小鼠IL15(mIL15)可包含SEQ ID NO：4所示的氨基酸序列。又例如，人IL15(hIL15)可包含SEQ ID NO：12所示的氨基酸序列。
在本申请中，术语“IL7”通常是指白细胞介素-7。IL7主要由胸腺和骨髓基质细胞分泌，属于相对分子质量为25000-28000的糖蛋白。IL7及其受体介导的信号转导途径主要是通过Janus激酶、信号转导及转录活化子和磷酸肌醇3-激酶三条通路实现的。例如，小鼠IL7(mIL7)可包含SEQ ID NO：5所示的氨基酸序列。又例如，人IL7(hIL7)可包含SEQ ID NO：13所示的氨基酸序列。
在本申请中，术语“IL21”通常是指白细胞介素-21。IL21由活化的CD4+T细胞分泌；IL21参与调节B细胞增殖，缺失IL21基因可导致机体更容易受细菌或病毒的感染。研究表明，IL21可通过调控Bcl-6蛋白影响IL2受体蛋白(CD25)的表达水平。例如，小鼠IL21(mIL21)可包含SEQ ID NO：6所示的氨基酸序列。又例如，人IL21(hIL21)可包含SEQ ID NO：14所示的氨基酸序列。
在本申请中，术语“FTL3L”通常是指FMS相关酪氨酸激酶3配体。FTL3L可调节非红系造血干细胞的增殖和分化，促进前B淋巴细胞、树突状细胞、NK细胞、细胞毒T淋巴细胞的增殖、分化和成熟，具有重要的抗肿瘤作用。例如，小鼠FTL3L(mFTL3L)可包含SEQ ID NO：7所示的氨基酸序列。又例如，人FTL3L(hFTL3L)可包含SEQ ID NO：15所示的氨基酸序列。
在本申请中，术语“GMCSF”通常是指粒细胞巨噬细胞集落刺激因子。GMCSF能刺激粒细胞和巨噬细胞的增殖、分化及活化，增加造血功能；亦能增强中性粒细胞、嗜酸性细胞及单核粒细胞的多种功能。GMCSF可提高免疫效应细胞吞噬细菌及杀灭癌细胞等免疫活力，并且有利于恢复因肿瘤化疗和骨髓移植引起的中性粒细胞缺乏。例如，小鼠GMCSF(mGMCSF)可包含SEQ ID NO：8所示的氨基酸序列。又例如，人GMCSF(hGMCSF)可包含SEQ ID NO：16所示的氨基酸序列。
在本申请中，术语“核酸分子”通常是指从其天然环境中分离的或人工合成的任何长度的分离形式的核苷酸、脱氧核糖核苷酸或核糖核苷酸或其类似物。在本申请中，所述核酸分子可编码所述的蛋白质异二聚体，其包括SEQ ID NO.38-57中任一项所示的核苷酸序列。
在本申请中，术语“载体”通常是指能够在合适的宿主中自我复制的核酸分子。所述载体可以将插入的核酸分子转移到宿主细胞中和/或宿主细胞之间。所述载体可包括主要用于将DNA或RNA插入细胞中的载体、主要用于复制DNA或RNA的载体，以及主要用于DNA或RNA的转录和/或翻译的表达的载体。所述载体还包括具有多种上述功能的载体。所述载体可以是当引入合适的宿主细胞时能够转录并翻译成多肽的多核苷酸。通常，通过培养包含所述载体的合适的宿主细胞，所述载体可以产生期望的表达产物。
在本申请中，术语“宿主细胞”通常是指可以或已经含有包括本申请所述的核酸分子的质粒或载体，或者能够表达本申请所述的蛋白质异二聚体的个体细胞、细胞系或细胞培养物。所述细胞可以包括单个宿主细胞的子代。由于天然的，意外的或故意的突变，子代细胞与原始亲本细胞在形态上或在基因组上可能不一定完全相同，但能够表达本申请所述的蛋白质异二聚体组合即可。所述宿主细胞可以通过使用本申请所述的载体体外转染该宿主细胞而得到。在本申请中，通过使用表达载体的病毒转染B16(rtTA)肿瘤细胞或293A细胞，可以得到能够表达本申请所述的蛋白质异二聚体的宿主细胞。
在本申请中，术语“药学上可接受的载体”通常是指药物组合物的组分之一，其可以包括缓冲剂、抗氧化剂、防腐剂、低分子量多肽、蛋白质、亲水聚合物、氨基酸、糖、螯合剂、反离子、金属复合物和/或非离子表面活性剂等。例如，所述药学上可接受的载体可以包括赋形剂，例如，所述赋形剂可以选自下组：淀粉、糊精、蔗糖、乳糖、硬脂酸镁、硫酸钙、羧甲基素、滑石粉、海藻酸钙凝胶、壳聚糖和纳米微球等。例如，所述药学上可接受的载体还可以选自下组：pH调节剂、渗透压调节剂、增溶剂和抑菌剂。
在本申请中，术语“肿瘤”通常是指或描述哺乳动物的生理状况，其典型特征在于细胞增殖或存活失调。例如，肿瘤可以选自以下组：肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
蛋白质异二聚体
一方面，本申请提供一种蛋白质异二聚体，其可以包含第一多肽链以及不同于所述第一多肽链的第二多肽链，其中所述第一多肽链可以包含IL12a以及与所述IL12a融合的第一因子，所述第二多肽链可以包含IL12b以及与所述IL12b融合的第二因子，且其中所述第一因子及所述第二因子可以各自独立地选自下组：IL2、GMCSF、IL7、IL15、IL21和FLT3L。
在本申请中，所述第一因子可以与所述第二因子不同。在本申请中，所述蛋白质异二聚体可以是一种细胞因子融合蛋白，其将所述细胞因子，即IL12和IL2、GMCSF、IL7、IL15、IL21和FLT3L中的两种或多种，通过基因重组技术融合在一起。所述蛋白质异二聚体可以既具有组成其因子独特的生物学活性，又可以通过生物学活性的互补及协同效应发挥出较单一细胞因子所不具备的生物学功能，甚至还可以产生一些新的结构及生物学功能。
在本申请中，所述第一因子和所述第二因子可以彼此不同地分别选自细胞因子组A和细胞因子组B，所述细胞因子组A可以选自下组：IL2、IL7、IL15和IL21，且细胞因子组B可以选自下组：GMCSF和FLT3L。
例如，第一因子可以选择细胞因子组A中的IL2，且第二因子可以选择细胞因子组B中的GMCSF；或第一因子可以选择细胞因子组A中的IL7，且第二因子可以选择细胞因子组B中的GMCSF；或第一因子可以选择细胞因子组A中的IL15，且第二因子可以选择细胞因子组B中的GMCSF；或第一因子可以选择细胞因子组A中的IL21，且第二因子可以选择细胞因子组B中的GMCSF；或第一因子可以选择细胞因子组A中的IL2，且第二因子可以选择细胞因子组B中的FLT3L；或第一因子可以选择细胞因子组A中的IL7，且第二因子可以选择细胞因子组B中的FLT3L；或第一因子可以选择细胞因子组A中的IL15，且第二因子可以选择细胞因子组B中的FLT3L；或第一因子可以选择细胞因子组A中的IL21，且第二因子可以选择细胞因子组B中的FLT3L；或第二因子可以选择细胞因子组A中的IL2，且第一因子可以选择细胞因子组B中的GMCSF；或第二因子可以选择细胞因子组A中的IL7，且第一因子可以选择细胞因子组B中的GMCSF；或第二因子可以选择细胞因子组A中的IL15，且第一因子可以选择细胞因子组B中的GMCSF；或第二因子可以选择细胞因子组A中的IL21，且第一因子可以选择细胞因子组B中的GMCSF；或第二因子可以选择细胞因子组A中的IL2，且第一因子可以选择细胞因子组B中的FLT3L；或第二因子可以选择细胞因子组A中的IL7，且第一因子可以选择细胞因子组B中的FLT3L；或第二因子可以选择细胞因子组A中的IL15，且第一因子可以选择细胞因子组B中的FLT3L；或第二因子可以选择细胞因子组A中的IL21，且第一因子可以选择细胞因子组B中的FLT3L。
在本申请中，在所述第一多肽链中，所述IL12a的C末端可以与所述第一因子的N末端直接或间接融合。例如，在所述第一多肽链中，所述IL12a的C末端可以与IL2的N端直接或间接融合；或在所述第一多肽链中，所述IL12a的C末端可以与IL7的N端直接或间接融合；或在所述第一多肽链中，所述IL12a的C末端可以与IL15的N端直接或间接融合；或在所述第一多肽链中，所述IL12a的C末端可以与IL21的N端直接或间接融合；或在所述第一多肽链中，所述IL12a的C末端可以与GMCSF的N端直接或间接融合；或在所述第一多肽链中，所述IL12a的C末端可以与FLT3L的N端直接或间接融合。
在本申请中，在所述第一多肽链中，所述IL12a的N末端可以与所述第一因子的C末端直接或间接融合。例如，在所述第一多肽链中，所述IL12a的N末端可以与IL2的C端直接或间接融合；或在所述第一多肽链中，所述IL12a的N末端可以与IL7的C端直接或间接融合；或在所述第一多肽链中，所述IL12a的N末端可以与IL15的C端直接或间接融合；或在所述第一多肽链中，所述IL12a的N末端可以与IL21的C端直接或间接融合；或在所述第一多肽链中，所述IL12a的N末端可以与GMCSF的C端直接或间接融合；或在所述第一多肽链中，所述IL12a的N末端可以与FLT3L的C端直接或间接融合。
在本申请中，在所述第二多肽链中，所述IL12b的C末端可以与所述第二因子的N末端直接或间接融合。例如，在所述第二多肽链中，所述IL12b的C末端可以与IL2的N端直接或间接融合；或在所述第二多肽链中，所述IL12b的C末端可以与IL7的N端直接或间接融合；或在所述第二多肽链中，所述IL12b的C末端可以与IL15的N端直接或间接融合；或在所述第二多肽链中，所述IL12b的C末端可以与IL21的N端直接或间接融合；或在所述第二多肽链中，所述IL12b的C末端可以与GMCSF的N端直接或间接融合；或在所述第二多肽链中，所述IL12b的C末端可以与FLT3L的N端直接或间接融合。
在本申请中，在所述第二多肽链中，所述IL12b的N末端可以与所述第二因子的C末端直接或间接融合。例如，在所述第二多肽链中，所述IL12b的N末端可以与IL2的C端直接或间接融合；或在所述第二多肽链中，所述IL12b的N末端可以与IL7的C端直接或间接融合；或在所述第二多肽链中，所述IL12b的N末端可以与IL15的C端直接或间接融合；或在所述第二多肽链中，所述IL12b的N末端可以与IL21的C端直接或间接融合；或在所述第二多肽链中，所述IL12b的N末端可以与GMCSF的C端直接或间接融合；或在所述第二多肽链中，所述IL12b的N末端可以与FLT3L的C端直接或间接融合。
在本申请中，所述蛋白质异二聚体可以包含以上所有所述细胞因子的位置连接关系的任意的排列组合。
在本申请中，所述第一因子和所述第二因子可以选自下组：
1)第一因子可以为IL2，且第二因子可以为GMCSF；
2)第一因子可以为IL7，且第二因子可以为GMCSF；
3)第一因子可以为IL15，且第二因子可以为GMCSF；
4)第一因子可以为IL21，且第二因子可以为GMCSF；
5)第一因子可以为IL2，且第二因子可以为FLT3L；
6)第一因子可以为IL7，且第二因子可以为FLT3L；
7)第一因子可以为IL15，且第二因子可以为FLT3L；
8)第一因子可以为IL21，且第二因子可以为FLT3L；
9)第一因子可以为GMCSF，且第二因子可以为IL2；
10)第一因子可以为GMCSF，且第二因子可以为IL7；
11)第一因子可以为GMCSF，且第二因子可以为IL15；
12)第一因子可以为GMCSF，且第二因子可以为IL21；
13)第一因子可以为FLT3L，且第二因子可以为IL2；
14)第一因子可以为FLT3L，且第二因子可以为IL7；
15)第一因子可以为FLT3L，且第二因子可以为IL15；以及，
16)第一因子可以为FLT3L，且第二因子可以为IL21。
在本申请中，所述IL12a和所述第一因子之间，和/或所述IL12b和所述第二因子之间可通过连接肽连接。例如，所述连接肽的氨基酸序列可以如SEQ ID.17所示。
在本申请中，所述IL12a和所述IL12b可以通过二硫键共价结合。在本申请中，所述第一多肽链和所述第二多肽链之间可以通过IL12a和IL12b之间的二硫键结合形成所述蛋白质异二聚体。在某些实施方式中，编码所述第一多肽链和所述第二多肽链的核苷酸序列之间可以包含编码自切割肽的核苷酸序列。例如，当包含上述核苷酸序列的表达载体转录并完成翻译后，所述自切割肽的位置会被切开，从而构成包括所述第一多肽链和所述第二多肽链且通过二硫键形成的所述蛋白质异二聚体。在某些实施方式中，构建表达载体时，所述第一多肽链和所述第二多肽链之间可以通过2A自剪切肽或柔性氨基酸接头肽连接。例如，所述2A自剪切肽的氨基酸序列可以如SEQ ID NO.58所示，编码所述2A自剪切肽的核苷酸序列可以如SEQ ID NO.59所示。又例如，所述柔性氨基酸接头肽的氨基酸序列可以如SEQID.17所示。
某些实施方式中，所述第一多肽链和所述第二多肽链之间也可以包含所述连接肽。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL2的N端融合形成第一多肽链，且IL12b的C端可以和GMCSF的N端融合形成第二多肽链，从而形成IL12a-IL2-IL12b-GMCSF蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL2的C端可以和IL12a的N端融合形成第一多肽链，且IL12b的C端可以和GMCSF的N端融合形成第二多肽链，从而形成IL2-IL12a-IL12b-GMCSF蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL2的N端融合形成第一多肽链，且GMCSF的C端可以和IL12b的N端融合形成第二多肽链，从而形成IL12a-IL2-GMCSF-IL12b蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL2的C端可以和IL12a的N端融合形成第一多肽链，且GMCSF的C端可以和IL12b的N端融合形成第二多肽链，从而形成IL2-IL12a-GMCSF-IL12b蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和GMCSF的N端融合形成第一多肽链，且IL12b的C端可以和IL2的N端融合形成第二多肽链，从而形成IL12a-GMCSF-IL12b-IL2蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL7的N端融合形成第一多肽链，且IL12b的C端可以和GMCSF的N端融合形成第二多肽链，从而形成IL12a-IL7-IL12b-GMCSF蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL15的N端融合形成第一多肽链，且IL12b的C端可以和GMCSF的N端融合形成第二多肽链，从而形成IL12a-IL15-IL12b-GMCSF蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL21的N端融合形成第一多肽链，且IL12b的C端可以和GMCSF的N端融合形成第二多肽链，从而形成IL12a-IL21-IL12b-GMCSF蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL2的N端融合形成第一多肽链，且IL12b的C端可以和FLT3L的N端融合形成第二多肽链，从而形成IL12a-IL2-IL12b-FLT3L蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL7的N端融合形成第一多肽链，且IL12b的C端可以和FLT3L的N端融合形成第二多肽链，从而形成IL12a-IL7-IL12b-FLT3L蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL15的N端融合形成第一多肽链，且IL12b的C端可以和FLT3L的N端融合形成第二多肽链，从而形成IL12a-IL15-IL12b-FLT3L蛋白质异二聚体。
例如，在所述蛋白质异二聚体中，IL12a的C端可以和IL21的N端融合形成第一多肽链，且IL12b的C端可以和FLT3L的N端融合形成第二多肽链，从而形成IL12a-IL21-IL12b-FLT3L蛋白质异二聚体。
在本申请中，所述IL12a、所述IL12b、所述第一因子和所述第二因子可以来源自哺乳动物。例如，所述哺乳动物可以选自下组：人、小鼠、大鼠、猴、犬、猪、羊、牛和猫。在本申请中，所述哺乳动物可以选自下组：人、小鼠。例如，所述IL12a、IL12b、IL2、GMCSF、IL7、IL15、IL21和FLT3L可以选自小鼠，所述小鼠来源的IL12a、IL12b、IL2、GMCSF、IL7、IL15、IL21和FLT3L可以简写作mIL12a、mIL12b、mIL2、mGMCSF、mIL7、mIL15、mIL21和mFLT3L。例如，所述IL12a、IL12b、IL2、GMCSF、IL7、IL15、IL21和FLT3L可以选自人，所述人来源的IL12a、IL12b、IL2、GMCSF、IL7、IL15、IL21和FLT3L可以简写作hIL12a、hIL12b、hIL2、hGMCSF、hIL7、hIL15、hIL21和hFLT3L。
在本申请中，所述IL12a、所述IL12b、所述第一因子和所述第二因子的来源可以相同。
在本申请中，所述第一多肽链可以包括如SEQ ID NO.60-65和SEQ ID NO.70-73中任一项所示的序列。
在本申请中，所述第二多肽链可以包括如SEQ ID NO.66-69和SEQ ID NO.74-75中任一项所示的序列。
在本申请中，所述蛋白质异二聚体可以包括如SEQ ID NO.18-37中任一项所示的序列。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL2的N端可以融合形成mIL12a-mIL2第一多肽链(序列如SEQ ID NO.60所示)，且mIL12b的C端和mGMCSF的N端可以融合形成mIL12b-mGMCSF第二多肽链(序列如SEQ ID NO.66所示)，可以形成mIL12a-mIL2-mIL12b-mGMCSF蛋白质异二聚体(序列如SEQ ID NO.18所示)。
例如，在所述蛋白质异二聚体中，mIL2的C端和mIL12a的N端可以融合形成mIL2-mIL12a第一多肽链(序列如SEQ ID NO.61所示)，且mIL12b的C端和mGMCSF的N端可以融合形成mIL12b-mGMCSF第二多肽链(序列如SEQ ID NO.66所示)，可以形成mIL2-mIL12a-mIL12b-mGMCSF蛋白质异二聚体(序列如SEQ ID NO.19所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL2的N端可以融合形成mIL12a-mIL2第一多肽链(序列如SEQ ID NO.60所示)，且mGMCSF的C端和mIL12b的N端可以融合形成mGMCSF-mIL12b第二多肽链(序列如SEQ ID NO.67所示)，可以形成mIL12a-mIL2-mGMCSF-mIL12b蛋白质异二聚体(序列如SEQ ID NO.20所示)。
例如，在所述蛋白质异二聚体中，mIL2的C端和mIL12a的N端可以融合形成mIL2-mIL12a第一多肽链(序列如SEQ ID NO.61所示)，且mGMCSF的C端和mIL12b的N端可以融合形成mGMCSF-mIL12b第二多肽链(序列如SEQ ID NO.67所示)，可以形成mIL2-mIL12a-mGMCSF-mIL12b蛋白质异二聚体(序列如SEQ ID NO.21所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mGMCSF的N端可以融合形成mIL12a-mGMCSF第一多肽链(序列如SEQ ID NO.62所示)，且mIL12b的C端和mIL2的N端可以融合形成mIL12b-mIL2第二多肽链(序列如SEQ ID NO.68所示)，可以形成mIL12a-mGMCSF-mIL12b-mIL2蛋白质异二聚体(序列如SEQ ID NO.22所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL7的N端可以融合形成mIL12a-mIL7第一多肽链(序列如SEQ ID NO.63所示)，且mIL12b的C端和mGMCSF的N端可以融合形成mIL12b-mGMCSF第二多肽链(序列如SEQ ID NO.66所示)，可以形成mIL12a-mIL7-mIL12b-mGMCSF蛋白质异二聚体(序列如SEQ ID NO.23所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL15的N端可以融合形成mIL12a-mIL15第一多肽链(序列如SEQ ID NO.64所示)，且mIL12b的C端和mGMCSF的N端可以融合形成mIL12b-mGMCSF第二多肽链(序列如SEQ ID NO.66所示)，可以形成mIL12a-mIL15-mIL12b-mGMCSF蛋白质异二聚体(序列如SEQ ID NO.24所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL21的N端可以融合形成mIL12a-mIL21第一多肽链(序列如SEQ ID NO.65所示)，且mIL12b的C端和mGMCSF的N端可以融合形成mIL12b-mGMCSF第二多肽链(序列如SEQ ID NO.66所示)，可以形成mIL12a-mIL21-mIL12b-mGMCSF蛋白质异二聚体(序列如SEQ ID NO.25所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL2的N端可以融合形成mIL12a-mIL2第一多肽链(序列如SEQ ID NO.60所示)，且mIL12b的C端和mFLT3L的N端可以融合形成mIL12b-mFLT3L第二多肽链(序列如SEQ ID NO.69所示)，可以形成mIL12a-mIL2-mIL12b-mFLT3L蛋白质异二聚体(序列如SEQ ID NO.26所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL7的N端可以融合形成mIL12a-mIL7第一多肽链(序列如SEQ ID NO.63所示)，且mIL12b的C端和mFLT3L的N端可以融合形成mIL12b-mFLT3L第二多肽链(序列如SEQ ID NO.69所示)，可以形成mIL12a-mIL7-mIL12b-mFLT3L蛋白质异二聚体(序列如SEQ ID NO.27所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL15的N端可以融合形成mIL12a-mIL15第一多肽链(序列如SEQ ID NO.64所示)，且mIL12b的C端和mFLT3L的N端可以融合形成mIL12b-mFLT3L第二多肽链(序列如SEQ ID NO.69所示)，可以形成mIL12a-mIL15-mIL12b-mFLT3L蛋白质异二聚体(序列如SEQ ID NO.28所示)。
例如，在所述蛋白质异二聚体中，mIL12a的C端和mIL21的N端可以融合形成mIL12a-mIL21第一多肽链(序列如SEQ ID NO.65所示)，且mIL12b的C端和mFLT3L的N端可以融合形成mIL12b-mFLT3L第二多肽链(序列如SEQ ID NO.69所示)，可以形成mIL12a-mIL21-mIL12b-mFLT3L蛋白质异二聚体(序列如SEQ ID NO.29所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL2的N端可以融合形成hIL12a-hIL2第一多肽链(序列如SEQ ID NO.70所示)，且hIL12b的C端和hGMCSF的N端可以融合形成hIL12b-hGMCSF第二多肽链(序列如SEQ ID NO.74所示)，可以形成hIL12a-hIL2-hIL12b-hGMCSF蛋白质异二聚体(序列如SEQ ID NO.30所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL7的N端可以融合形成hIL12a-hIL7第一多肽链(序列如SEQ ID NO.71所示)，且hIL12b的C端和hGMCSF的N端可以融合形成hIL12b-hGMCSF第二多肽链(序列如SEQ ID NO.74所示)，可以形成hIL12a-hIL7-hIL12b-hGMCSF蛋白质异二聚体(序列如SEQ ID NO.31所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL15的N端可以融合形成hIL12a-hIL15第一多肽链(序列如SEQ ID NO.72所示)，且hIL12b的C端和hGMCSF的N端可以融合形成hIL12b-hGMCSF第二多肽链(序列如SEQ ID NO.74所示)，可以形成hIL12a-hIL15-hIL12b-hGMCSF蛋白质异二聚体(序列如SEQ ID NO.32所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL21的N端可以融合形成hIL12a-hIL21第一多肽链(序列如SEQ ID NO.73所示)，且hIL12b的C端和hGMCSF的N端可以融合形成hIL12b-hGMCSF第二多肽链(序列如SEQ ID NO.74所示)，可以形成hIL12a-hIL21-hIL12b-hGMCSF蛋白质异二聚体(序列如SEQ ID NO.33所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL2的N端可以融合形成hIL12a-hIL2第一多肽链(序列如SEQ ID NO.70所示)，且hIL12b的C端和hFLT3L的N端可以融合形成hIL12b-hFLT3L第二多肽链(序列如SEQ ID NO.75所示)，可以形成hIL12a-hIL2-hIL12b-hFLT3L蛋白质异二聚体(序列如SEQ ID NO.34所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL7的N端可以融合形成hIL12a-hIL7第一多肽链(序列如SEQ ID NO.71所示)，且hIL12b的C端和hFLT3L的N端可以融合形成hIL12b-hFLT3L第二多肽链(序列如SEQ ID NO.75所示)，可以形成hIL12a-hIL7-hIL12b-hFLT3L蛋白质异二聚体(序列如SEQ ID NO.35所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL15的N端可以融合形成hIL12a-hIL15第一多肽链(序列如SEQ ID NO.72所示)，且hIL12b的C端和hFLT3L的N端可以融合形成hIL12b-hFLT3L第二多肽链(序列如SEQ ID NO.75所示)，可以形成hIL12a-hIL15-hIL12b-hFLT3L蛋白质异二聚体(序列如SEQ ID NO.36所示)。
例如，在所述蛋白质异二聚体中，hIL12a的C端和hIL21的N端可以融合形成hIL12a-hIL21第一多肽链(序列如SEQ ID NO.73所示)，且hIL12b的C端和hFLT3L的N端可以融合形成hIL12b-hFLT3L第二多肽链(序列如SEQ ID NO.75所示)，可以形成hIL12a-hIL21-hIL12b-hFLT3L蛋白质异二聚体(序列如SEQ ID NO.37所示)。
在本申请中，所述mIL12a-mIL2-mIL12b-mGMCSF、mIL2-mIL12a-mIL12b-mGMCSF、mIL12a-mIL2-mGMCSF-mIL12b、mIL2-mIL12a-mGMCSF-mIL12b、mIL12a-mGMCSF-mIL12b-mIL2、mIL12a-mIL7-mIL12b-mGMCSF、mIL12a-mIL15-mIL12b-mGMCSF、mIL12a-mIL21-mIL12b-mGMCSF、mIL12a-mIL2-mIL12b-mFLT3L、mIL12a-mIL7-mIL12b-mFLT3L、mIL12a-mIL15-mIL12b-mFLT3L、mIL12a-mIL21-mIL12b-mFLT3L、hIL12a-hIL2-hIL12b-hGMCSF、hIL12a-hIL7-hIL12b-hGMCSF、hIL12a-hIL15-hIL12b-hGMCSF、hIL12a-hIL21-hIL12b-hGMCSF、hIL12a-hIL2-hIL12b-hFLT3L、hIL12a-hIL7-hIL12b-hFLT3L、hIL12a-hIL15-hIL12b-hFLT3L和hIL12a-hIL21-hIL12b-hFLT3L可以分别依次简称为mIL12aIL2IL12bGMCSF、mIL2IL12aIL12bGMCSF、mIL12aIL2GMCSFIL12b、mIL2IL12aGMCSFIL12b、mIL12aGMCSFIL12bIL2、mIL12aIL7IL12bGMCSF、mIL12aIL15IL12bGMCSF、mIL12aIL21IL12bGMCSF、mIL12aIL2IL12bFLT3L、mIL12aIL7IL12bFLT3L、mIL12aIL15IL12bFLT3L、mIL12aIL21IL12bFLT3L、hIL12aIL2IL12bGMCSF、hIL12aIL7IL12bGMCSF、hIL12aIL15IL12bGMCSF、hIL12aIL21IL12bGMCSF、hIL12aIL2IL12bFLT3L、hIL12aIL7IL12bFLT3L、hIL12aIL15IL12bFLT3L和hIL12aIL21IL12bFLT3L。上述蛋白质异二聚体的结构可以如图2所示。
在本申请中，所述的蛋白质异二聚体可以用于治疗肿瘤。
另一方面，本申请提供了一种所述的蛋白质异二聚体在制备药物中的用途，所述药物可以用于治疗肿瘤。
所述肿瘤可以选自以下组：黑色素瘤、肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
核酸分子、表达载体、宿主细胞、药物组合物及制备方法
另一方面，本申请提供了一种或多种分离的核酸分子，其可以编码所述的蛋白质异二聚体。例如，所述核酸分子可以包括如SEQ ID NO.38-57中任一项所示的序列。例如，编码所述mIL12aIL2IL12bGMCSF、mIL2IL12aIL12bGMCSF、mIL12aIL2GMCSFIL12b、mIL2IL12aGMCSFIL12b、mIL12aGMCSFIL12bIL2、mIL12aIL7IL12bGMCSF、mIL12aIL15IL12bGMCSF、mIL12aIL21IL12bGMCSF、mIL12aIL2IL12bFLT3L、mIL12aIL7IL12bFLT3L、mIL12aIL15IL12bFLT3L、mIL12aIL21IL12bFLT3L、hIL12aIL2IL12bGMCSF、hIL12aIL7IL12bGMCSF、hIL12aIL15IL12bGMCSF、hIL12aIL21IL12bGMCSF、hIL12aIL2IL12bFLT3L、hIL12aIL7IL12bFLT3L、hIL12aIL15IL12bFLT3L和hIL12aIL21IL12bFLT3L的核苷酸序列可以分别为SEQ ID NO.38、SEQ ID NO.39、SEQ ID NO.40、SEQ ID NO.41、SEQ ID NO.42、SEQ ID NO.43、SEQ IDNO.44、SEQ ID NO.45、SEQ ID NO.46、SEQ ID NO.47、SEQ ID NO.48、SEQ ID NO.49、SEQ IDNO.50、SEQ ID NO.51、SEQ ID NO.52、SEQ ID NO.53、SEQ ID NO.54、SEQ ID NO.55、SEQ IDNO.56和SEQ ID NO.57。
本申请中，所述核酸分子可以用于治疗肿瘤。
另一方面，本申请提供了一种所述核酸分子在制备药物中的用途，所述药物可以用于治疗肿瘤。
所述肿瘤可以选自以下组：黑色素瘤、肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
另一方面，本申请提供了一种表达载体，其可以包括编码所述的蛋白质异二聚体的核苷酸序列。在本申请中，可以通过分别将编码mIL12aIL2IL12bGMCSF、mIL2IL12aIL12bGMCSF、mIL12aIL2GMCSFIL12b、mIL2IL12aGMCSFIL12b、mIL12aGMCSFIL12bIL2、mIL12aIL7IL12bGMCSF、mIL12aIL15IL12bGMCSF、mIL12aIL21IL12bGMCSF、mIL12aIL2IL12bFLT3L、mIL12aIL7IL12bFLT3L、mIL12aIL15IL12bFLT3L和mIL12aIL21IL12bFLT3L的核苷酸序列插入到pLentis-PTRE-MCS-PGK-PURO核苷酸序列中，和，从而构建可调控表达载体pLentis-PTRE-mIL12aIL2IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL2IL12aIL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL2GMCSFIL12b-PGK-PURO、pLentis-PTRE-mIL2IL12aGMCSFIL12b-PGK-PURO、pLentis-PTRE-mIL12aGMCSFIL12bIL2-PGK-PURO、pLentis-PTRE-mIL12aIL7IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL15IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL21IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL2IL12bFLT3L-PGK-PURO、pLentis-PTRE-mIL12aIL7IL12bFLT3L-PGK-PURO、pLentis-PTRE-mIL12aIL15IL12bFLT3L-PGK-PURO和pLentis-PTRE-mIL12aIL21IL12bFLT3L-PGK-PURO。例如，可以通过分别将hIL12aIL2IL12bGMCSF、hIL12aIL7IL12bGMCSF、hIL12aIL15IL12bGMCSF、hIL12aIL21IL12bGMCSF、hIL12aIL2IL12bFLT3L、hIL12aIL7IL12bFLT3L、hIL12aIL15IL12bFLT3L和hIL12aIL21IL12bFLT3L插入到pLentis-CMV-MCS-IRES-PURO中，从而构建表达载体pLentis-CMV-hIL12aIL2IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL7IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL15IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL21IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL2IL12bFLT3L-IRES-PURO、pLentis-CMV-hIL12aIL7IL12bFLT3L-IRES-PURO、pLentis-CMV-hIL12aIL15IL12bFLT3L-IRES-PURO和pLentis-CMV-hIL12aIL21IL12bFLT3L-IRES-PURO。
本申请中，所述表达载体可以用于治疗肿瘤。
另一方面，本申请提供了一种所述表达载体在制备药物中的用途，所述药物可以用于治疗肿瘤。所述肿瘤可以选自以下组：黑色素瘤、肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
在本申请中，构建载体和质粒的方法，例如将编码蛋白的基因插入到载体和质粒的方法或将质粒引入宿主细胞的方法，对于本领域中具有普通技术的人员是众所周知的，并且在许多出版物中都有所描述，包括Sambrook，J.，Fritsch，E.F.and Maniais，T.(1989)Molecular Cloning：A Laboratory Manual，2nd edition，Cold spring HarborLaboratory Press。
另一方面，本中请提供了一种宿主细胞，其可以包括所述的表达载体。在本申请中，所述宿主细胞可以是使用所述表达载体转染B16(rtTA)肿瘤细胞制备而得到的。例如，可以分别使用pLentis-PTRE-mIL12aIL2IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL2IL12aIL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL2GMCSFIL12b-PGK-PURO、pLentis-PTRE-mIL2IL12aGMCSFIL12b-PGK-PURO、pLentis-PTRE-mIL12aGMCSFIL12bIL2-PGK-PURO、pLentis-PTRE-mIL12aIL7IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL15IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL21IL12bGMCSF-PGK-PURO、pLentis-PTRE-mIL12aIL2IL12bFLT3L-PGK-PURO、pLentis-PTRE-mIL12aIL7IL12bFLT3L-PGK-PURO、pLentis-PTRE-mIL12aIL15IL12bFLT3L-PGK-PURO和pLentis-PTRE-mIL12aIL21IL12bFLT3L-PGK-PURO表达载体的病毒转染B16(rtTA)肿瘤细胞，从而得到B16(rtTA)-mIL12aIL2IL12bGMCSF、B16(rtTA)-mIL2IL12aIL12bGMCSF、B16(rtTA)-mIL12aIL2GMCSFIL12b、B16(rtTA)-mIL2IL12aGMCSFIL12b、B16(rtTA)-mIL12aGMCSFIL12bIL2、B16(rtTA)-mIL12aIL7IL12bGMCSF、B16(rtTA)-mIL12aIL15IL12bGMCSF、B16(rtTA)-mIL12aIL21IL12bGMCSF、B16(rtTA)-mIL12aIL2IL12bFLT3L、B16(rtTA)-mIL12aIL7IL12bFLT3L、B16(rtTA)-mIL12aIL15IL12bFLT3L和B16(rtTA)-mIL12aIL21IL12bFLT3L宿主细胞。例如，可以分别使用pLentis-CMV-hIL12aIL2IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL7IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL15IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL21IL12bGMCSF-IRES-PURO、pLentis-CMV-hIL12aIL2IL12bFLT3L-IRES-PURO、pLentis-CMV-hIL12aIL7IL12bFLT3L-IRES-PURO和pLentis-CMV-hIL12aIL15IL12bFLT3L-IRES-PURO、pLentis-CMV-hIL12aIL21IL12bFLT3L-IRES-PURO表达载体的病毒转染293A细胞，从而得到293A-hIL12aIL2IL12bGMCSF、293A-hIL12aIL7IL12bGMCSF、293A-hIL12aIL15IL12bGMCSF、293A-hIL12aIL21IL12bGMCSF、293A-hIL12aIL2IL12bFLT3L、293A-hIL12aIL7IL12bFLT3L、293A-hIL12aIL15IL12bFLT3L和293A-hIL12aIL21IL12bFLT3L宿主细胞。
本申请中，所述宿主细胞可以用于治疗肿瘤。所述肿瘤可以选自以下组：黑色素瘤、肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
另一方面，本申请提供了一种所述宿主细胞在制备药物中的用途，所述药物可以用于治疗肿瘤。所述肿瘤可以选自以下组：黑色素瘤、肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
本申请提供了蛋白质异二聚体、核酸分子、表达载体、宿主细胞、药物组合物，其用于治疗肿瘤。
治疗肿瘤的方法，其包括以下的步骤：向有需要的受试者施用本申请所述的蛋白质异二聚体、核酸分子、表达载体、宿主细胞和/或药物组合物。
所述肿瘤可以选自以下组：黑色素瘤、肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
另一方面，本申请提供了一种制备所述的蛋白质异二聚体的方法，其可以包括以下的步骤：在能够表达所述的蛋白质异二聚体的情况下，培养所述的宿主细胞。
药物组合物及其用途
另一方面，本申请提供了一种药物组合物，其可以包括所述的蛋白质异二聚体、所述的核酸分子、所述的载体和/或所述的宿主细胞，以及药学上可接受的载体。
所述药学上可接受的载体可以包括缓冲剂、抗氧化剂、防腐剂、低分子量多肽、蛋白质、亲水聚合物、氨基酸、糖、螯合剂、反离子、金属复合物和/或非离子表面活性剂等。例如，所述药学上可接受的载体可以包括赋形剂，例如，所述赋形剂可以选自下组：淀粉、糊精、蔗糖、乳糖、硬脂酸镁、硫酸钙、羧甲基素、滑石粉、海藻酸钙凝胶、壳聚糖和纳米微球等。例如，所述药学上可接受的载体还可以选自下组：pH调节剂、渗透压调节剂、增溶剂和抑菌剂。
在本申请中，所述药物组合物可被配制用于口服给药，静脉内给药，肌肉内给药，在肿瘤部位的原位给药，吸入，直肠给药，阴道给药，经皮给药或通过皮下储存库给药。
在本申请中，所述药物组合物可以用于治疗肿瘤。所述肿瘤可以选自以下组：黑色素瘤、肺癌、食道癌、胃癌、结直肠癌、肝癌、乳腺癌、宫颈癌、甲状腺癌、脑及中枢神经系统癌、胰腺癌、口腔癌、鼻咽癌、头颈癌、喉癌、骨癌、皮肤癌、卵巢癌、前列腺癌、睾丸癌、肾癌、膀胱癌、眼睑肿瘤、白血病和淋巴瘤。
在本申请中，所述药物组合物可以用于抑制肿瘤生长。例如，本申请的药物组合物可以抑制或延缓疾病的发展或进展，可以通过促进细胞因子的表达减小肿瘤的大小(甚至基本消除肿瘤)，和/或可以减轻和/或稳定疾病状态。
在本申请中，所述药物组合物可以含有治疗有效量的所述的蛋白质异二聚体、所述的核酸分子、所述的载体和/或所述的宿主细胞。所述治疗有效量是能够预防和/或治疗(至少部分治疗)患有或具有发展风险的受试者中的病症或病症(例如癌症)和/或其任何并发症而所需的剂量。
另一方面，本申请提供一种缓解或治疗肿瘤的方法，所述方法可包括施用所述蛋白质异二聚体、所述的核酸分子、所述的表达载体、所述宿主细胞和/或所述药物组合物。
在本申请中，所述施用的方法可包括口服给药，静脉内给药，肌肉内给药，在肿瘤部位的原位给药，吸入，直肠给药，阴道给药，经皮给药或通过皮下储存库给药。
下面显示的实施例意在说明本发明的具体实施方案，并且不意在以任何方式限制本说明书或权利要求书的范围。
实施例
试剂：DMEM培养基、1640培养基、胎牛血清购自lifetechnologies公司；细胞培养瓶、培养板购自Corning公司；强力霉素(DOX)购自上海生工生物工程有限公司；嘌呤霉素(Puromycin)、Blasticidin购自Chemicon公司；限制性内切酶购自Takara和NEB公司；连接酶购自NEB公司；DNA聚合酶购自Takara公司；质粒提取试剂盒、胶回收试剂盒购自OmegaBiotech公司；引物合成由上海生工生物工程有限公司完成；基因合成由南京金斯瑞公司完成；ELISA试剂盒购于博士德公司。细胞因子mIL12、mGMCSF、mFLT3L、mIL2、mIL15、mIL21、mIL7、hIL12、hGMCSF、hFLT3L、hIL2、hIL15、hIL21和hIL7购于Peprotech公司。
实施例的部分试验结果参见图1、4-6。在图1和图4-图16中，每条折线各代表一只小鼠体内肿瘤的面积状况，例如，5条折线表示一次实验的5只小鼠的数据；分数则表示肿瘤清除的小鼠/接种肿瘤的小鼠的比例，如10/10表示肿瘤清除的小鼠/接种肿瘤的小鼠是10/10＝100％。
实施例1表达可调控蛋白的肿瘤细胞的制备
1.1构建第一表达载体pLentis-CMV-rtTA-IRES-Bsd
合成两端带有BamHI和EcoRI位点的rtTA(GenBank：ALK44378.1)的DNA序列，合成产物连接在载体pUC57上。酶切所述连接有rtTA的pUC57载体，酶切体系如下：6μg连接有rtTA的pUC57载体质粒、4μl酶切缓冲液、1μl BamHI和1μl EcoRI，加水至总体积为40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收rtTA片段，待用。
在EP管内酶切载体pLentis-CMV-IRES-Bsd，酶切体系如下：2μg pLentis-CMV-IRES-Bsd载体质粒、3μl酶切缓冲液、1μl BamHI和1μl EcoRI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-CMV-IRES-Bsd载体片段，待用。
连接pLentis-CMV-IRES-Bsd和rtTA，体系如下：2μl pLentis-CMV-IRES-Bsd、2μlrtTA、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水，置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中，37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定rtTA片段是否成功连入pLentis-CMV-IRES-Bsd载体中，然后将正确的载体送测序，确定第一表达载体pLentis-CMV-rtTA-IRES-Bsd构建成功。
1.2制备第一表达载体pLentis-CMV-rtTA-IRES-Bsd的病毒
1)消化培养的293FT细胞，计数后将3×106细胞/孔铺入10cm培养皿中，培养液体积为10ml。
2)第二天晚上，观察细胞状态，如果细胞状态合适良好，进行转染。在培养板中加入氯喹至终浓度25μM，取一只试管，加入灭菌水及以下质粒(5μg pMD2G+15μg pSPAX2+20μgpLentis-CMV-rtTA-IRES-Bsd)，总体积为1045μl，然后加入155μl 2MCaCl2，混匀，最后再加入1200μl 2×HBS，边滴加边振荡，滴加完毕后，迅速将混合物加入到细胞培养孔中，轻轻摇晃混匀。
3)第三天早上，观察细胞状态，将培养基换为10ml新鲜DMEM培养基。
4)第五天早上，观察细胞状态，并收集培养皿中的上清，用0.45μm滤器过滤，然后置于高速离心管中，50000g离心2小时，小心弃去上清，尽量用吸水纸吸干液体，然后用500μlHBSS重悬沉淀，溶解2小时后分装成小管，-70℃保存，得到第一表达载体pLentis-CMV-rtTA-IRES-Bsd的病毒。
1.3使用第一表达载体pLentis-CMV-rtTA-IRES-Bsd的病毒转染B16肿瘤细胞
消化培养的小鼠黑色素瘤细胞B16，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl第一表达载体pLentis-CMV-rtTA-IRES-Bsd的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按适合此细胞的浓度加入杀稻瘟菌素(blasticidin)，继续培养，每两天更换一次培养基，并保持杀稻瘟菌素的浓度为8μg/ml，筛选一周后，存活的细胞即为稳定表达调控蛋白的细胞，将此细胞命名为B16(rtTA)。
实施例2诱导表达绿色荧光蛋白(GFP)对肿瘤生长的影响
2.1构建编码绿色荧光蛋白(GFP)的可调控表达载体
使用引物，以带有GFP基因为模板进行PCR反应扩增GFP基因，PCR条件按PrimeStarHSDNApolymerase说明书进行。PCR进行琼脂糖凝胶电泳后，用胶回收试剂盒回收，然后用BamHI和EcoRI进行酶切，酶切体系如下：30μg PCR回收产物、4μl酶切缓冲液、1μlBamHI和1μl EcoRI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收GFP基因片段，待用。
酶切可调控表达载体，体系如下：2μg pLentis-PTRE-MCS-PGK-PURO质粒、3μl酶切缓冲液、1μl BamHI和1μl EcoRI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收载体片段，待用。连接pLentis-PTRE-MCS-PGK-PURO和GFP，连接体系如下：2μl pLentis-PTRE-MCS-PGK-PURO、2μl GFP、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-GFP-PGK-PURO构建成功。
2.2制备可调控表达GFP的细胞
制备GFP表达载体的病毒，方法与第一表达载体的病毒制备方法一致，得到第二表达载体pLentis-PTRE-GFP-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-GFP-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素(puromycin)，继续培养三天后，存活的细胞即为可调控表达GFP的细胞，此细胞命名为B16(rtTA)-GFP。
2.3调控表达GFP对肿瘤生长的影响
将对数生长期的B16(rtTA)-GFP细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠(购于北京华阜康生物科技股份有限公司)的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况，如图1所示，结果显示诱导表达GFP对肿瘤生长无抑制作用(其中每条折线各代表一只小鼠体内肿瘤的面积状况)。
实施例3设计蛋白质异二聚体
设计包括如下蛋白质异二聚体：mIL2IL12aIL12bGMCSF、mIL12aIL2GMCSFIL12b、mIL2IL12aGMCSFIL12b、mIL12aGMCSFIL12bIL2、mIL12aIL7IL12bGMCSF、mIL12aIL15IL12bGMCSF、mIL12aIL21IL12bGMCSF、mIL12aIL2IL12bFLT3L、mIL12aIL7IL12bFLT3L、mIL12aIL15IL12bFLT3L、mIL12aIL21IL12bFLT3L、hIL12aIL2IL12bGMCSF、hIL12aIL7IL12bGMCSF、hIL12aIL15IL12bGMCSF、hIL12aIL21IL12bGMCSF、hIL12aIL2IL12bFLT3L、hIL12aIL7IL12bFLT3L、hIL12aIL15IL12bFLT3L和hIL12aIL21IL12bFLT3L。上述蛋白质异二聚体的结构如图2所示。
实施例4诱导表达mIL12aIL2IL12bGMCSF对肿瘤生长的影响
4.1构建mIL12aIL2IL12bGMCSF可调控表达载体
合成mIL12aIL2IL12bGMCSF编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL2IL12bGMCSF质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL2IL12bGMCSF基因片段，待用。
mIL12aIL2IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.18所示，编码所述mIL12aIL2IL12bGMCSF的核苷酸序列如SEQ ID NO.38所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL2IL12bGMCSF，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL2IL12bGMCSF、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL2IL12bGMCSF-PGK-PURO构建成功。
4.2制备可调控表达mIL12aIL2IL12bGMCSF的细胞
制备所述mIL12aIL2IL12bGMCSF表达载体的病毒，方法与第一表达载体的病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL2IL12bGMCSF-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-mIL12aIL2IL12bGMCSF-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL2IL12bGMCSF的细胞，此细胞命名为B16(rtTA)-mIL12aIL2IL12bGMCSF。
4.3体外检测mIL12aIL2IL12bGMCSF的诱导表达效果
将B16(rtTA)-mIL12aIL2IL12bGMCSF细胞按5×104每孔铺入24孔板中，24小时后，在孔内加入100ng/ml的强力霉素(DOX)，继续培养72小时，收集上清。使用小鼠IL12p70ELISA试剂盒检测上清中的融合蛋白表达量，操作按试剂盒说明书进行。未加DOX诱导的孔作为对照。如图3所示，DOX的加入诱导了mIL12aIL2IL12bGMCSF融合蛋白的表达。
4.4诱导表达mIL12aIL2IL12bGMCSF对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL2IL12bGMCSF细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝10/10，如图4所示，mIL12aIL2IL12bGMCSF能够在全部小鼠中诱导肿瘤消退。
实施例5诱导表达mIL2IL12aIL12bGMCSF对肿瘤生长的影响
5.1构建mIL2IL12aIL12bGMCSF可调控表达载体
合成mIL2IL12aIL12bGMCSFF基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL2IL12aIL12bGMCSF质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL2IL12aIL12bGMCSF基因片段，待用。
mIL2IL12aIL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO 19所示，编码所述mIL2IL12aIL12bGMCSF的核苷酸序列是SEQ ID NO.39。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL2IL12aIL12bGMCSF，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL2IL12aIL12bGMCSF、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL2IL12aIL12bGMCSF-PGK-PURO构建成功。
5.2制备可调控表达mIL2IL12aIL12bGMCSF的细胞
制备所述mIL2IL12aIL12bGMCSF表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL2IL12aIL12bGMCSF-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二表达载体pLentis-PTRE-mIL2IL12aIL12bGMCSF-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL2IL12aIL12bGMCSF的细胞，此细胞命名为B16(rtTA)-mIL2IL12aIL12bGMCSF。
5.3诱导表达mIL2IL12aIL12bGMCSF对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL2IL12aIL12bGMCSF细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝10/10，如图5所示，mIL2IL12aIL12bGMCSF能够在全部小鼠中诱导肿瘤消退。
实施例6诱导表达mIL12aIL2GMCSFIL12b对肿瘤生长的影响
6.1构建mIL12aIL2GMCSFIL12b可调控表达载体
合成mIL12aIL2GMCSFIL12b基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL2GMCSFIL12b质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL2GMCSFIL12b基因片段，待用。
mIL12aIL2GMCSFIL12b蛋白质异二聚体的氨基酸序列如SEQ ID NO.20所示，编码所述mIL12aIL2GMCSFIL12b的核苷酸序列如SEQ ID NO.40所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μgpLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL2GMCSFIL12b，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL2GMCSFIL12b、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL2GMCSFIL12b-PGK-PURO构建成功。
6.2制备可调控表达mIL12aIL2GMCSFIL12b的细胞
制备所述mIL12aIL2GMCSFIL12b表达载体的病毒，方法与第一表达载体的病毒制备方法一致，得到第二可调控表达载体pLentis-PTRE-mIL12aIL2GMCSFIL12b-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-mIL12aIL2GMCSFIL12b-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL2GMCSFIL12b的细胞，此细胞命名为B16(rtTA)-mIL12aIL2GMCSFIL12b。
6.3诱导表达mIL12aIL2GMCSFIL12b对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL2GMCSFIL12b细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝10/10，如图6所示，mIL12aIL2GMCSFIL12b能够在全部小鼠中诱导肿瘤消退。
实施例7诱导表达mIL2IL12aGMCSFIL12b对肿瘤生长的影响
7.1构建mIL2IL12aGMCSFIL12b可调控表达载体
合成mIL2IL12aGMCSFIL12b基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL2IL12aGMCSFIL12b质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL2IL12aGMCSFIL12b基因片段，待用。
mIL2IL12aGMCSFIL12b蛋白质异二聚体的氨基酸序列如SEQ ID NO.21所示，编码所述mIL2IL12aGMCSFIL12b的核苷酸序列如SEQ ID NO.41所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO质粒、3μl酶切缓冲液、1μl BamHI、1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL2IL12aGMCSFIL12b，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL2IL12aGMCSFIL12b、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL2IL12aGMCSFIL12b-PGK-PURO构建成功。
7.2制备可调控表达mIL2IL12aGMCSFIL12b的细胞
制备所述mIL2IL12aGMCSFIL12b表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL2IL12aGMCSFIL12b-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二表达调控载体pLentis-PTRE-mIL2IL12aGMCSFIL12b-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL2IL12aGMCSFIL12b的细胞，此细胞命名为B16(rtTA)-mIL2IL12aGMCSFIL12b。
7.3诱导表达mIL2IL12aGMCSFIL12b对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL2IL12aGMCSFIL12b细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝10/10，如图7所示，mIL2IL12aGMCSFIL12b能够在全部小鼠中诱导肿瘤消退。
实施例8诱导表达mIL12aGMCSFIL12bIL2对肿瘤生长的影响
8.1构建mIL12aGMCSFIL12bIL2可调控表达载体
合成mIL12aGMCSFIL12bIL2基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aGMCSFIL12bIL2质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aGMCSFIL12bIL2基因片段，待用。
mIL12aGMCSFIL12bIL2蛋白质异二聚体的氨基酸序列如SEQ ID NO.22所示，编码所述mIL12aGMCSFIL12bIL2的核苷酸序列是SEQ ID NO.42。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aGMCSFIL12bIL2，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aGMCSFIL12bIL2、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aGMCSFIL12bIL2-PGK-PURO构建成功。
8.2制备可调控表达mIL12aGMCSFIL12bIL2的细胞
制备所述mIL12aGMCSFIL12bIL2表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aGMCSFIL12bIL2-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-mIL12aGMCSFIL12bIL2-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aGMCSFIL12bIL2的细胞，此细胞命名为B16(rtTA)-mIL12aGMCSFIL12bIL2。
8.3诱导表达mIL12aGMCSFIL12bIL2对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aGMCSFIL12bIL2细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝10/10，如图8所示，结果显示mIL12aGMCSFIL12bIL2能够在全部小鼠中诱导肿瘤消退。
实施例9诱导表达hIL12aIL2IL12bGMCSF对肿瘤生长的影响
9.1构建hIL12aIL2IL12bGMCSF可调控表达载体
合成hIL12aIL2IL12bGMCSF基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μghIL12aIL2IL12bGMCSF质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收hIL12aIL2IL12bGMCSF基因片段，待用。
hIL12aIL2IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.30所示，编码所述hIL12aIL2IL12bGMCSF的核苷酸序列如SEQ ID NO.50所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和hIL12aIL2IL12bGMCSF，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl hIL12aIL2IL12bGMCSF、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-hIL12aIL2IL12bGMCSF-PGK-PURO构建成功。
9.2制备可调控表达hIL12aIL2IL12bGMCSF的细胞
制备所述hIL12aIL2IL12bGMCSF表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-hIL12aIL2IL12bGMCSF-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二表达载体pLentis-PTRE-hIL12aIL2IL12bGMCSF-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达hIL12aIL2IL12bGMCSF的细胞，此细胞命名为B16(rtTA)-hIL12aIL2IL12bGMCSF。
9.3诱导表达hIL12aIL2IL12bGMCSF对肿瘤生长的影响
将对数生长期的B16(rtTA)-hIL12aIL2IL12bGMCSF细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，如图9所示，hIL12aIL2IL12bGMCSF能够一定程度上抑制小鼠肿瘤的生长。
实施例10诱导表达mIL12aIL7IL12bGMCSF对肿瘤生长的影响
10.1构建mIL12aIL7IL12bGMCSF可调控表达载体
合成mIL12aIL7IL12bGMCSF基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL7IL12bGMCSF质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL7IL12bGMCSF基因片段，待用。
mIL12aIL7IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.23所示，编码所述mIL12aIL7IL12bGMCSF的核苷酸序列如SEQ ID NO.43所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI、1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL7IL12bGMCSF，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL7IL12bGMCSF、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL7IL12bGMCSF-PGK-PURO构建成功。
10.2制备可调控表达mIL12aIL7IL12bGMCSF的细胞
制备所述mIL12aIL7IL12bGMCSF表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL7IL12bGMCSF-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-mIL12aIL7IL12bGMCSF-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL7IL12bGMCSF的细胞，此细胞命名为B16(rtTA)-mIL12aIL7IL12bGMCSF。
10.3诱导表达mIL12aIL7IL12bGMCSF对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL7IL12bGMCSF细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝10/10，如图10所示，mIL12aIL7IL12bGMCSF能够在全部小鼠中诱导肿瘤消退。
实施例11诱导表达mIL12aIL15IL12bGMCSF对肿瘤生长的影响
11.1构建mIL12aIL15IL12bGMCSF可调控表达载体
合成mIL12aIL15IL12bGMCSF基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL15IL12bGMCSF质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL15IL12bGMCSF基因片段，待用。
mIL12aIL15IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.24所示，编码所述mIL12aIL15IL12bGMCSF的核苷酸序列如SEQ ID NO.44所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL15IL12bGMCSF，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL15IL12bGMCSF、1μl连接酶缓冲液、0.5μlT4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL15IL12bGMCSF-PGK-PURO构建成功。
11.2制备可调控表达mIL12aIL15IL12bGMCSF的细胞
制备所述mIL12aIL15IL12bGMCSF表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL15IL12bGMCSF-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-mIL12aIL15IL12bGMCSF-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL15IL12bGMCSF的细胞，此细胞命名为B16(rtTA)-mIL12aIL15IL12bGMCSF。
11.3诱导表达mIL12aIL15IL12bGMCSF对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL15IL12bGMCSF细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝8/10，如图11所示，mIL12aIL15IL12bGMCSF能够在部分小鼠中诱导肿瘤消退。
实施例12诱导表达mIL12aIL21IL12bGMCSF对肿瘤生长的影响
12.1构建mIL12aIL21IL12bGMCSF可调控表达载体
合成mIL12aIL21IL12bGMCSF基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL21IL12bGMCSF质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL21IL12bGMCSF基因片段，待用。
mIL12aIL21IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.25所示，编码所述mIL12aIL21IL12bGMCSF的核苷酸序列如SEQ ID NO.45所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI、1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL21IL12bGMCSF，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL21IL12bGMCSF、1μl连接酶缓冲液、0.5μlT4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL21IL12bGMCSF-PGK-PURO构建成功。
12.2制备可调控表达mIL12aIL21IL12bGMCSF的细胞
制备所述mIL12aIL21IL12bGMCSF表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL21IL12bGMCSF-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二表达载体pLentis-PTRE-mIL12aIL21IL12bGMCSF-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL21IL12bGMCSF的细胞，此细胞命名为B16(rtTA)-mIL12aIL21IL12bGMCSF。
12.3诱导表达mIL12aIL21IL12bGMCSF对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL21IL12bGMCSF细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝10/10，如图12所示，mIL12aIL21IL12bGMCSF能够在全部小鼠中诱导肿瘤消退。
实施例13诱导表达mIL12aIL2IL12bFLT3L对肿瘤生长的影响
13.1构建mIL12aIL2IL12bFLT3L可调控表达载体
合成mIL12aIL2IL12bFLT3L基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL2IL12bFLT3L质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL2IL12bFLT3L基因片段，待用。
mIL12aIL2IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.26所示，编码所述mIL12aIL2IL12bFLT3L的核苷酸序列如SEQ ID NO.46所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μg pLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL2IL12bFLT3L，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL2IL12bFLT3L、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL2IL12bFLT3L-PGK-PURO构建成功。
13.2制备可调控表达mIL12aIL2IL12bFLT3L的细胞
制备所述mIL12aIL2IL12bFLT3L表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL2IL12bFLT3L-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-mLL12aIL2IL12bFLT3L-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL2IL12bFLT3L的细胞，此细胞命名为B16(rtTA)-mIL12aIL2IL12bFLT3L。
13.3诱导表达mIL12aIL2IL12bFLT3L对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL2IL12bFLT3L细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝9/10，如图13所示，mIL12aIL2IL12bFLT3L能够在部分小鼠中诱导肿瘤消退。
实施例14诱导表达mIL12aIL7IL12bFLT3L对肿瘤生长的影响
14.1构建mIL12aIL7IL12bFLT3L可调控表达载体
合成mIL12aIL7IL12bFLT3L基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL7IL12bFLT3L质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL7IL12bFLT3L基因片段，待用。
mIL12aIL7IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.27所示，编码所述mIL12aIL7IL12bFLT3L的核苷酸序列如SEQ ID NO.47所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μgpLentis-PTRE-MCS-PGK-PURO质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL7IL12bFLT3L，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL7IL12bFLT3L、1μl连接酶缓冲液、0.5μl T4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL7IL12bFLT3L-PGK-PURO构建成功。
14.2制备可调控表达mIL12aIL7IL12bFLT3L的细胞
制备所述mIL12aIL7IL12bFLT3L表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL7IL12bFLT3L-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二表达载体pLentis-PTRE-mIL12aIL7IL12bFLT3L-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL7IL12bFLT3L的细胞，此细胞命名为B16(rtTA)-mIL12aIL7IL12bFLT3L。
14.3诱导表达mIL12aIL7IL12bFLT3L对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL7IL12bFLT3L细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝9/10，如图14所示，mIL12aIL7IL12bFLT3L能够在部分小鼠中诱导肿瘤消退。
实施例15诱导表达mIL12aIL15IL12bFLT3L对肿瘤生长的影响
15.1构建mIL12aIL15IL12bFLT3L可调控表达载体
合成mIL12aIL15IL12bFLT3L基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL15IL12bFLT3L质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL15IL12bFLT3L基因片段，待用。
mIL12aIL15IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.28所示，编码所述mIL12aIL15IL12bFLT3L的核苷酸序列如SEQ ID NO.48所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μgpLentis-PTRE-MCS-PGK-PURO载体质粒、3μl酶切缓冲液、1μl BamHI、1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL15IL12bFLT3L，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL15IL12bFLT3L、1μl连接酶缓冲液、0.5μlT4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL15IL12bFLT3L-PGK-PURO构建成功。
15.2制备可调控表达mIL12aIL15IL12bFLT3L的细胞
制备所述mIL12aIL15IL12bFLT3L表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL15IL12bFLT3L-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二表达载体pLentis-PTRE-mIL12aIL15IL12bFLT3L-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL15IL12bFLT3L的细胞，此细胞命名为B16(rtTA)-mIL12aIL15IL12bFLT3L。
15.3诱导表达mIL12aIL15IL12bFLT3L对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL15IL12bFLT3L细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝7/10，如图15所示，mIL12aIL15IL12bFLT3L能够在部分小鼠中诱导肿瘤消退。
实施例16诱导表达mIL12aIL21IL12bFLT3L对肿瘤生长的影响
16.1构建mIL12aIL21IL12bFLT3L可调控表达载体
合成mIL12aIL21IL12bFLT3L基因编码序列，两端分别带有BamHI、BglII和XhoI、EcoRI酶切位点，然后用BamHI或BglII和XhoI或EcoRI进行酶切，酶切体系如下：5μgmIL12aIL21IL12bFLT3L质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收mIL12aIL21IL12bFLT3L基因片段，待用。
mIL12aIL21IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.29所示，编码所述mIL12aIL21IL12bFLT3L的核苷酸序列如SEQ ID NO.49所示。
酶切可调控表达载体pLentis-PTRE-MCS-PGK-PURO，酶切体系如下：2μgpLentis-PTRE-MCS-PGK-PURO质粒、3μl酶切缓冲液、1μl BamHI、1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-PTRE-MCS-PGK-PURO载体片段，待用。
连接pLentis-PTRE-MCS-PGK-PURO和mIL12aIL21IL12bFLT3L，连接体系如下：2μlpLentis-PTRE-MCS-PGK-PURO、2μl mIL12aIL21IL12bFLT3L、1μl连接酶缓冲液、0.5μlT4DNA连接酶和4.5μl水。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37℃摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体送测序，确定第二表达载体pLentis-PTRE-mIL12aIL21IL12bFLT3L-PGK-PURO构建成功。
16.2制备可调控表达mIL12aIL21IL12bFLT3L的细胞
制备所述mIL12aIL21IL12bFLT3L表达载体的病毒，方法与第一表达载体病毒制备方法一致，得到第二表达载体pLentis-PTRE-mIL12aIL21IL12bFLT3L-PGK-PURO的病毒。
消化培养的B16(rtTA)肿瘤细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml，24小时后，加入10μl上述第二可调控表达载体pLentis-PTRE-mIL12aIL21IL12bFLT3L-PGK-PURO的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养，待细胞长满后，将其传出到培养瓶中，按终浓度3μg/ml加入嘌呤霉素，继续培养三天后，存活的细胞即为可调控表达mIL12aIL21IL12bFLT3L的细胞，此细胞命名为B16(rtTA)-mIL12aIL21IL12bFLT3L。
16.3诱导表达mIL12aIL21IL12bFLT3L对肿瘤生长的影响
将对数生长期的B16(rtTA)-mIL12aIL21IL12bFLT3L细胞消化，用HBSS稀释到2×106个/ml，使用1ml注射器按50μl/只注射到8-10周龄的C57BL/6雌性小鼠的右背侧，共10只，待肿瘤长出后使用含2g/L强力霉素的水喂养，记录小鼠肿瘤生长情况以及肿瘤清除率，肿瘤清除小鼠数/总的小鼠数＝9/10，如图16所示，mIL12aIL21IL12bFLT3L能够在部分小鼠中诱导肿瘤消退。
实施例17构建表达人hIL12aIL2IL12bGMCSF、人hIL12aIL7IL12bGMCSF、人hIL12aIL15IL12bGMCSF、人hIL12aIL21IL12bGMCSF、人hIL12aIL2IL12bFLT3L、人hIL12aIL7IL12bFLT3L、人hIL12aIL15IL12bFLT3L、人hIL12aIL21IL12bFLT3L的细胞
17.1构建可调控表达目的基因的载体
在EP管内酶切载体pLentis-CMV-MCS-IRES-PURO，体系如下：2μg pLentis-CMV-MCS-IRES-PURO载体质粒、3μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积30μl，37℃静置12小时。取出EP管，加入3.3μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收pLentis-CMV-MCS-IRES-PURO载体片段，待用。
分别合成人hIL12aIL2IL12bGMCSF、人hIL12aIL7IL12bGMCSF、人hIL12aIL15IL12bGMCSF、人hIL12aIL21IL12bGMCSF、人hIL12aIL2IL12bFLT3L、人hIL12aIL7IL12bFLT3L、人hIL12aIL15IL12bFLT3L和人hIL12aIL21IL12bFLT3L的DNA序列，合成时在其5’端加上BamHI或BglII酶切位点，在其3’端加上XhoI或EcoRI酶切位点。酶切合成的带有目的基因的质粒，体系如下：5μg质粒、4μl酶切缓冲液、1μl BamHI和1μl XhoI，加水至总体积40μl，37℃静置12小时。取出EP管，加入4.4μl 10×上样缓冲液，用1％琼脂糖凝胶进行电泳，电泳后回收片段，待用。
hIL12aIL2IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.30所示，编码所述hIL12aIL2IL12bGMCSF的核苷酸序列如SEQ ID NO.50所示。
hIL12aIL7IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.31所示，编码所述hIL12aIL7IL12bGMCSF的核苷酸序列如SEQ ID NO.51所示。
hIL12aIL15IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.32所示，编码所述hIL12aIL15IL12bGMCSF的核苷酸序列如SEQ ID NO.52所示。
hIL12aIL21IL12bGMCSF蛋白质异二聚体的氨基酸序列如SEQ ID NO.33所示，编码所述hIL12aIL21IL12bGMCSF的核苷酸序列如SEQ ID NO.53所示。
hIL12aIL2IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.34所示，编码所述hIL12aIL2IL12bFLT3L的核苷酸序列如SEQ ID NO.54所示。
hIL12aIL7IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.35所示，编码所述hIL12aIL7IL12bFLT3L的核苷酸序列如SEQ ID NO.55所示。
hIL12aIL15IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.36所示，编码所述hIL12aIL15IL12bFLT3L的核苷酸序列如SEQ ID NO.56所示。
hIL12aIL21IL12bFLT3L蛋白质异二聚体的氨基酸序列如SEQ ID NO.37所示，编码所述hIL12aIL21IL12bFLT3L的核苷酸序列如SEQ ID NO.57所示。
分别连接pLentis-CMV-MCS-IRES-PURO和人hIL12aIL2IL12bGMCSF、人hIL12aIL7IL12bGMCSF、人hIL12aIL15IL12bGMCSF、人hIL12aIL21IL12bGMCSF、人hIL12aIL2IL12bFLT3L、人hIL12aIL7IL12bFLT3L、人hIL12aIL15IL12bFLT3L和人hIL12aIL21IL12bFLT3L，体系如下：2μl pLentis-CMV-MCS-IRES-PURO载体片段、2μl基因片段、1μl连接酶缓冲液、0.5μl T4 DNA连接酶和水4.5μl。置于室温连接4小时。然后将连接体系进行大肠杆菌感受态的转化。第二天从转化的平板上挑取菌落，置于LB培养基中37度摇床内过夜培养，使用质粒提取试剂盒从培养的细菌中提取质粒，通过酶切鉴定片段是否成功连入载体中，然后将正确的载体测序，确定构建成功。获得表达目的基因的载体pLentis-CMV-hIL12aIL2IL12bGMCSF-IRES-PURO、载体pLentis-CMV-hIL12aIL7IL12bGMCSF-IRES-PURO、载体pLentis-CMV-hIL12aIL15IL12bGMCSF-IRES-PURO、载体pLentis-CMV-hIL12aIL21IL12bGMCSF-IRES-PURO、载体pLentis-CMV-hIL12aIL2IL12bFLT3L-IRES-PURO、载体pLentis-CMV-hIL12aIL7IL12bFLT3L-IRES-PURO、载体pLentis-CMV-hIL12aIL15IL12bFLT3L-IRES-PURO和载体pLentis-CMV-hIL12aIL21IL12bFLT3L-IRES-PURO。
17.2制备可调控表达目的基因的细胞
1)消化培养的293FT细胞，计数后将3×106个细胞/孔铺入10cm培养皿中，培养液体积为10ml，共铺5块板。
2)第二天晚上，观察细胞状态，如果细胞状态好，进行转染。在培养板中加入氯喹至终浓度25μM，取一只试管，加入灭菌水及以下质粒(pMD2.G 6μg+pSPAX215μg+表达载体20μg)，总体积为1045μl，然后加入2M CaCl2155μl，混匀，最后再加入1200μl 2×HBS，边滴加边振荡，滴加完毕后，迅速将混合物加入到细胞培养孔中，轻轻摇晃混匀。
3)第三天早上，观察细胞状态，将培养基换为10ml新鲜DMEM培养基。
4)第五天早上，观察细胞状态，并收集培养皿中的上清，用0.45μm滤器过滤，然后置于高速离心管中，50000g离心2小时，小心弃去上清，尽量用吸水纸吸干液体，然后用200μl HBSS重悬沉淀，溶解2小时后分装成小管，-70℃保存。
17.3制备可调控表达目的基因的细胞
消化培养的293A细胞，按105个细胞/孔接种到6孔板中，培养体积为1ml。24小时后，加入10μl表达上述目的基因的病毒，在培养箱内继续培养24小时后，弃去上清，换为新鲜的培养基继续培养。待细胞长满后，将其传出到培养瓶中，加入终浓度3μg/ml嘌呤霉素，继续培养，每两天更换一次培养基，并保持嘌呤霉素的浓度，筛选一周后，存活的细胞即为稳定表达所述细胞因子的细胞，分别命名为293A-hIL12aIL2IL12bGMCSF、293A-hIL12aIL7IL12bGMCSF、293A-hIL12aIL15IL12bGMCSF、293A-hIL12aIL21IL12bGMCSF、293A-hIL12aIL2IL12bFLT3L、293A-hIL12aIL7IL12bFLT3L、293A-hIL12aIL15IL12bFLT3L、293A-hIL12aIL21IL12bFLT3L。
将构建的表达细胞，按5×104个每孔铺入到24孔板中，培养96小时后，收集上清，用人IL12p70ELISA试剂盒检测上清中融合蛋白的表达，操作按说明书进行。如图17所示，这些细胞均能够产生大量的IL12p70成功构建了表达融合蛋白的细胞。
前述详细说明是以解释和举例的方式提供的，并非要限制所附权利要求的范围。目前本文所列举的实施方式的多种变化对本领域普通技术人员来说是显而易见的，且保留在所附的权利要求和其等同方案的范围内。
序列表
<110> 张晋宇
<120> 蛋白质异二聚体及其用途
<130> 0069-PA-008CN
<160> 75
<170> PatentIn version 3.5
<210> 1
<211> 193
<212> PRT
<213> Mus musculus
<400> 1
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala
<210> 2
<211> 313
<212> PRT
<213> Mus musculus
<400> 2
Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr
1 5 10 15
Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys
85 90 95
Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser
100 105 110
Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys
115 120 125
Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr
130 135 140
Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg
145 150 155 160
Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro
165 170 175
Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln
180 185 190
Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile
195 200 205
Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn
210 215 220
Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro
225 230 235 240
His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys
245 250 255
Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe
260 265 270
Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val
275 280 285
Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp
290 295 300
Ala Cys Val Pro Cys Arg Val Arg Ser
305 310
<210> 3
<211> 149
<212> PRT
<213> Mus musculus
<400> 3
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
1 5 10 15
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
20 25 30
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
35 40 45
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
50 55 60
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
65 70 75 80
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
85 90 95
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
100 105 110
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
115 120 125
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
130 135 140
Ser Thr Ser Pro Gln
145
<210> 4
<211> 114
<212> PRT
<213> Mus musculus
<400> 4
Asn Trp Ile Asp Val Arg Tyr Asp Leu Glu Lys Ile Glu Ser Leu Ile
1 5 10 15
Gln Ser Ile His Ile Asp Thr Thr Leu Tyr Thr Asp Ser Asp Phe His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu Thr Val Arg
50 55 60
Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn Lys Asn Val
65 70 75 80
Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Thr Phe
85 90 95
Thr Glu Phe Leu Gln Ser Phe Ile Arg Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 5
<211> 129
<212> PRT
<213> Mus musculus
<400> 5
Glu Cys His Ile Lys Asp Lys Glu Gly Lys Ala Tyr Glu Ser Val Leu
1 5 10 15
Met Ile Ser Ile Asp Glu Leu Asp Lys Met Thr Gly Thr Asp Ser Asn
20 25 30
Cys Pro Asn Asn Glu Pro Asn Phe Phe Arg Lys His Val Cys Asp Asp
35 40 45
Thr Lys Glu Ala Ala Phe Leu Asn Arg Ala Ala Arg Lys Leu Lys Gln
50 55 60
Phe Leu Lys Met Asn Ile Ser Glu Glu Phe Asn Val His Leu Leu Thr
65 70 75 80
Val Ser Gln Gly Thr Gln Thr Leu Val Asn Cys Thr Ser Lys Glu Glu
85 90 95
Lys Asn Val Lys Glu Gln Lys Lys Asn Asp Ala Cys Phe Leu Lys Arg
100 105 110
Leu Leu Arg Glu Ile Lys Thr Cys Trp Asn Lys Ile Leu Lys Gly Ser
115 120 125
Ile
<210> 6
<211> 129
<212> PRT
<213> Mus musculus
<400> 6
His Lys Ser Ser Pro Gln Gly Pro Asp Arg Leu Leu Ile Arg Leu Arg
1 5 10 15
His Leu Ile Asp Ile Val Glu Gln Leu Lys Ile Tyr Glu Asn Asp Leu
20 25 30
Asp Pro Glu Leu Leu Ser Ala Pro Gln Asp Val Lys Gly His Cys Glu
35 40 45
His Ala Ala Phe Ala Cys Phe Gln Lys Ala Lys Leu Lys Pro Ser Asn
50 55 60
Pro Gly Asn Asn Lys Thr Phe Ile Ile Asp Leu Val Ala Gln Leu Arg
65 70 75 80
Arg Arg Leu Pro Ala Arg Arg Gly Gly Lys Lys Gln Lys His Ile Ala
85 90 95
Lys Cys Pro Ser Cys Asp Ser Tyr Glu Lys Arg Thr Pro Lys Glu Phe
100 105 110
Leu Glu Arg Leu Lys Trp Leu Leu Gln Lys Met Ile His Gln His Leu
115 120 125
Ser
<210> 7
<211> 206
<212> PRT
<213> Mus musculus
<400> 7
Gly Thr Pro Asp Cys Tyr Phe Ser His Ser Pro Ile Ser Ser Asn Phe
1 5 10 15
Lys Val Lys Phe Arg Glu Leu Thr Asp His Leu Leu Lys Asp Tyr Pro
20 25 30
Val Thr Val Ala Val Asn Leu Gln Asp Glu Lys His Cys Lys Ala Leu
35 40 45
Trp Ser Leu Phe Leu Ala Gln Arg Trp Ile Glu Gln Leu Lys Thr Val
50 55 60
Ala Gly Ser Lys Met Gln Thr Leu Leu Glu Asp Val Asn Thr Glu Ile
65 70 75 80
His Phe Val Thr Ser Cys Thr Phe Gln Pro Leu Pro Glu Cys Leu Arg
85 90 95
Phe Val Gln Thr Asn Ile Ser His Leu Leu Lys Asp Thr Cys Thr Gln
100 105 110
Leu Leu Ala Leu Lys Pro Cys Ile Gly Lys Ala Cys Gln Asn Phe Ser
115 120 125
Arg Cys Leu Glu Val Gln Cys Gln Pro Asp Ser Ser Thr Leu Leu Pro
130 135 140
Pro Arg Ser Pro Ile Ala Leu Glu Ala Thr Glu Leu Pro Glu Pro Arg
145 150 155 160
Pro Arg Gln Leu Leu Leu Leu Leu Leu Leu Leu Leu Pro Leu Thr Leu
165 170 175
Val Leu Leu Ala Ala Ala Trp Gly Leu Arg Trp Gln Arg Ala Arg Arg
180 185 190
Arg Gly Glu Leu His Pro Gly Val Pro Leu Pro Ser His Pro
195 200 205
<210> 8
<211> 124
<212> PRT
<213> Mus musculus
<400> 8
Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val
1 5 10 15
Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr
20 25 30
Leu Asn Glu Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys
35 40 45
Leu Thr Cys Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg
50 55 60
Gly Asn Phe Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr
65 70 75 80
Tyr Gln Thr Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln
85 90 95
Val Thr Thr Tyr Ala Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr
100 105 110
Asp Ile Pro Phe Glu Cys Lys Lys Pro Gly Gln Lys
115 120
<210> 9
<211> 197
<212> PRT
<213> Homo sapiens
<400> 9
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser
195
<210> 10
<211> 306
<212> PRT
<213> Homo sapiens
<400> 10
Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr
1 5 10 15
Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys
85 90 95
Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys
100 105 110
Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr
115 120 125
Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln
130 135 140
Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly
145 150 155 160
Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala
165 170 175
Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala
180 185 190
Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg
195 200 205
Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu
210 215 220
Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp
225 230 235 240
Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln
245 250 255
Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr
260 265 270
Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala
275 280 285
Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro
290 295 300
Cys Ser
305
<210> 11
<211> 133
<212> PRT
<213> Homo sapiens
<400> 11
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 12
<211> 114
<212> PRT
<213> Homo sapiens
<400> 12
Asn Trp Val Asn Val Ile Ser Asp Leu Lys Lys Ile Glu Asp Leu Ile
1 5 10 15
Gln Ser Met His Ile Asp Ala Thr Leu Tyr Thr Glu Ser Asp Val His
20 25 30
Pro Ser Cys Lys Val Thr Ala Met Lys Cys Phe Leu Leu Glu Leu Gln
35 40 45
Val Ile Ser Leu Glu Ser Gly Asp Ala Ser Ile His Asp Thr Val Glu
50 55 60
Asn Leu Ile Ile Leu Ala Asn Asn Ser Leu Ser Ser Asn Gly Asn Val
65 70 75 80
Thr Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu Lys Asn Ile
85 90 95
Lys Glu Phe Leu Gln Ser Phe Val His Ile Val Gln Met Phe Ile Asn
100 105 110
Thr Ser
<210> 13
<211> 152
<212> PRT
<213> Homo sapiens
<400> 13
Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr Glu Ser Val Leu
1 5 10 15
Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys Glu Ile Gly Ser
20 25 30
Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg His Ile Cys Asp
35 40 45
Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala Arg Lys Leu Arg
50 55 60
Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp Leu His Leu Leu
65 70 75 80
Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys Thr Gly Gln Val
85 90 95
Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln Pro Thr Lys Ser
100 105 110
Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys Leu Asn Asp Leu
115 120 125
Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr Cys Trp Asn Lys
130 135 140
Ile Leu Met Gly Thr Lys Glu His
145 150
<210> 14
<211> 133
<212> PRT
<213> Homo sapiens
<400> 14
Gln Gly Gln Asp Arg His Met Ile Arg Met Arg Gln Leu Ile Asp Ile
1 5 10 15
Val Asp Gln Leu Lys Asn Tyr Val Asn Asp Leu Val Pro Glu Phe Leu
20 25 30
Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu Trp Ser Ala Phe Ser
35 40 45
Cys Phe Gln Lys Ala Gln Leu Lys Ser Ala Asn Thr Gly Asn Asn Glu
50 55 60
Arg Ile Ile Asn Val Ser Ile Lys Lys Leu Lys Arg Lys Pro Pro Ser
65 70 75 80
Thr Asn Ala Gly Arg Arg Gln Lys His Arg Leu Thr Cys Pro Ser Cys
85 90 95
Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe Leu Glu Arg Phe Lys
100 105 110
Ser Leu Leu Gln Lys Met Ile His Gln His Leu Ser Ser Arg Thr His
115 120 125
Gly Ser Glu Asp Ser
130
<210> 15
<211> 209
<212> PRT
<213> Homo sapiens
<400> 15
Thr Gln Asp Cys Ser Phe Gln His Ser Pro Ile Ser Ser Asp Phe Ala
1 5 10 15
Val Lys Ile Arg Glu Leu Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val
20 25 30
Thr Val Ala Ser Asn Leu Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp
35 40 45
Arg Leu Val Leu Ala Gln Arg Trp Met Glu Arg Leu Lys Thr Val Ala
50 55 60
Gly Ser Lys Met Gln Gly Leu Leu Glu Arg Val Asn Thr Glu Ile His
65 70 75 80
Phe Val Thr Lys Cys Ala Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe
85 90 95
Val Gln Thr Asn Ile Ser Arg Leu Leu Gln Glu Thr Ser Glu Gln Leu
100 105 110
Val Ala Leu Lys Pro Trp Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu
115 120 125
Glu Leu Gln Cys Gln Pro Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser
130 135 140
Pro Arg Pro Leu Glu Ala Thr Ala Pro Thr Ala Pro Gln Pro Pro Leu
145 150 155 160
Leu Leu Leu Leu Leu Leu Pro Val Gly Leu Leu Leu Leu Ala Ala Ala
165 170 175
Trp Cys Leu His Trp Gln Arg Thr Arg Arg Arg Thr Pro Arg Pro Gly
180 185 190
Glu Gln Val Pro Pro Val Pro Ser Pro Gln Asp Leu Leu Leu Val Glu
195 200 205
His
<210> 16
<211> 127
<212> PRT
<213> Homo sapiens
<400> 16
Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gln Pro Trp Glu His Val
1 5 10 15
Asn Ala Ile Gln Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr
20 25 30
Ala Ala Glu Met Asn Glu Thr Val Glu Val Ile Ser Glu Met Phe Asp
35 40 45
Leu Gln Glu Pro Thr Cys Leu Gln Thr Arg Leu Glu Leu Tyr Lys Gln
50 55 60
Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met Met
65 70 75 80
Ala Ser His Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser Cys
85 90 95
Ala Thr Gln Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp
100 105 110
Phe Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu
115 120 125
<210> 17
<211> 15
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 连接肽
<400> 17
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 18
<211> 809
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL2IL12bGMCSF蛋白质异二聚体
<400> 18
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
210 215 220
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
225 230 235 240
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
245 250 255
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
260 265 270
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
275 280 285
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
290 295 300
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
305 310 315 320
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
325 330 335
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
340 345 350
Ser Thr Ser Pro Gln Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val
355 360 365
Glu Val Asp Trp Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr
370 375 380
Cys Asp Thr Pro Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg
385 390 395 400
His Gly Val Ile Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu
405 410 415
Phe Leu Asp Ala Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu
420 425 430
Ser His Ser His Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser
435 440 445
Thr Glu Ile Leu Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu
450 455 460
Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg
465 470 475 480
Asn Met Asp Leu Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp
485 490 495
Ser Arg Ala Val Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val
500 505 510
Thr Leu Asp Gln Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu
515 520 525
Asp Val Thr Cys Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala
530 535 540
Leu Glu Ala Arg Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe
545 550 555 560
Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met
565 570 575
Lys Pro Leu Lys Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp
580 585 590
Ser Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg
595 600 605
Ile Gln Arg Lys Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn
610 615 620
Gln Lys Gly Ala Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys
625 630 635 640
Lys Gly Gly Asn Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser
645 650 655
Ser Cys Ser Lys Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly
660 665 670
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr
675 680 685
Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val Glu Ala Ile
690 695 700
Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu
705 710 715 720
Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys
725 730 735
Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe
740 745 750
Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr
755 760 765
Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr
770 775 780
Tyr Ala Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro
785 790 795 800
Phe Glu Cys Lys Lys Pro Gly Gln Lys
805
<210> 19
<211> 809
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL2IL12aIL12bGMCSF蛋白质异二聚体
<400> 19
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
1 5 10 15
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
20 25 30
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
35 40 45
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
50 55 60
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
65 70 75 80
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
85 90 95
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
100 105 110
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
115 120 125
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
130 135 140
Ser Thr Ser Pro Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
145 150 155 160
Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu
165 170 175
Ser Gln Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr
180 185 190
Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp
195 200 205
His Glu Asp Ile Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu
210 215 220
Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr
225 230 235 240
Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu
245 250 255
Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr
260 265 270
Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His
275 280 285
Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu
290 295 300
Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro
305 310 315 320
Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu
325 330 335
Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly
340 345 350
Tyr Leu Ser Ser Ala Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val
355 360 365
Glu Val Asp Trp Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr
370 375 380
Cys Asp Thr Pro Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg
385 390 395 400
His Gly Val Ile Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu
405 410 415
Phe Leu Asp Ala Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu
420 425 430
Ser His Ser His Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser
435 440 445
Thr Glu Ile Leu Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu
450 455 460
Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg
465 470 475 480
Asn Met Asp Leu Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp
485 490 495
Ser Arg Ala Val Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val
500 505 510
Thr Leu Asp Gln Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu
515 520 525
Asp Val Thr Cys Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala
530 535 540
Leu Glu Ala Arg Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe
545 550 555 560
Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met
565 570 575
Lys Pro Leu Lys Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp
580 585 590
Ser Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg
595 600 605
Ile Gln Arg Lys Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn
610 615 620
Gln Lys Gly Ala Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys
625 630 635 640
Lys Gly Gly Asn Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser
645 650 655
Ser Cys Ser Lys Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly
660 665 670
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr
675 680 685
Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val Glu Ala Ile
690 695 700
Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu
705 710 715 720
Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys
725 730 735
Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe
740 745 750
Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr
755 760 765
Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr
770 775 780
Tyr Ala Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro
785 790 795 800
Phe Glu Cys Lys Lys Pro Gly Gln Lys
805
<210> 20
<211> 809
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL2GMCSFIL12b蛋白质异二聚体
<400> 20
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
210 215 220
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
225 230 235 240
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
245 250 255
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
260 265 270
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
275 280 285
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
290 295 300
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
305 310 315 320
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
325 330 335
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
340 345 350
Ser Thr Ser Pro Gln Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg
355 360 365
Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu Asp
370 375 380
Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val Ser Asn Glu
385 390 395 400
Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu Lys Ile Phe
405 410 415
Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly Ala Leu Asn
420 425 430
Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr Pro Glu Thr
435 440 445
Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile Asp Ser Leu
450 455 460
Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys Pro Gly Gln
465 470 475 480
Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
485 490 495
Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr
500 505 510
Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu
515 520 525
Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly
530 535 540
Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly
545 550 555 560
Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu
565 570 575
Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys
580 585 590
Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser
595 600 605
Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys
610 615 620
Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr
625 630 635 640
Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg
645 650 655
Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro
660 665 670
Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln
675 680 685
Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile
690 695 700
Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn
705 710 715 720
Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro
725 730 735
His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys
740 745 750
Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe
755 760 765
Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val
770 775 780
Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp
785 790 795 800
Ala Cys Val Pro Cys Arg Val Arg Ser
805
<210> 21
<211> 809
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL2IL12aGMCSFIL12b蛋白质异二聚体
<400> 21
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
1 5 10 15
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
20 25 30
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
35 40 45
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
50 55 60
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
65 70 75 80
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
85 90 95
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
100 105 110
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
115 120 125
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
130 135 140
Ser Thr Ser Pro Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
145 150 155 160
Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu
165 170 175
Ser Gln Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr
180 185 190
Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp
195 200 205
His Glu Asp Ile Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu
210 215 220
Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr
225 230 235 240
Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu
245 250 255
Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr
260 265 270
Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His
275 280 285
Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu
290 295 300
Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro
305 310 315 320
Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu
325 330 335
Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly
340 345 350
Tyr Leu Ser Ser Ala Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg
355 360 365
Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu Asp
370 375 380
Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val Ser Asn Glu
385 390 395 400
Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu Lys Ile Phe
405 410 415
Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly Ala Leu Asn
420 425 430
Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr Pro Glu Thr
435 440 445
Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile Asp Ser Leu
450 455 460
Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys Pro Gly Gln
465 470 475 480
Lys Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
485 490 495
Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr
500 505 510
Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu
515 520 525
Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly
530 535 540
Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly
545 550 555 560
Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu
565 570 575
Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys
580 585 590
Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser
595 600 605
Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys
610 615 620
Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr
625 630 635 640
Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg
645 650 655
Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro
660 665 670
Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln
675 680 685
Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile
690 695 700
Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn
705 710 715 720
Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro
725 730 735
His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys
740 745 750
Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe
755 760 765
Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val
770 775 780
Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp
785 790 795 800
Ala Cys Val Pro Cys Arg Val Arg Ser
805
<210> 22
<211> 809
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aGMCSFIL12bIL2蛋白质异二聚体
<400> 22
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val
210 215 220
Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr
225 230 235 240
Leu Asn Glu Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys
245 250 255
Leu Thr Cys Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg
260 265 270
Gly Asn Phe Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr
275 280 285
Tyr Gln Thr Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln
290 295 300
Val Thr Thr Tyr Ala Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr
305 310 315 320
Asp Ile Pro Phe Glu Cys Lys Lys Pro Gly Gln Lys Met Trp Glu Leu
325 330 335
Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr Pro Asp Ala Pro
340 345 350
Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu Glu Asp Asp Ile
355 360 365
Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly Ser Gly Lys Thr
370 375 380
Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly Gln Tyr Thr Cys
385 390 395 400
His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu Leu Leu His Lys
405 410 415
Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys Asn Phe Lys Asn
420 425 430
Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser Gly Arg Phe Thr
435 440 445
Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys Phe Asn Ile Lys
450 455 460
Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr Cys Gly Met Ala
465 470 475 480
Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg Asp Tyr Glu Lys
485 490 495
Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro Thr Ala Glu Glu
500 505 510
Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln Gln Asn Lys Tyr
515 520 525
Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp
530 535 540
Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn Ser Gln Val Glu
545 550 555 560
Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro His Ser Tyr Phe
565 570 575
Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys Glu Lys Met Lys
580 585 590
Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe Leu Val Glu Lys
595 600 605
Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val Cys Val Gln Ala
610 615 620
Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp Ala Cys Val Pro
625 630 635 640
Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
645 650 655
Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala
660 665 670
Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu
675 680 685
Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn
690 695 700
Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu
705 710 715 720
Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu
725 730 735
Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe
740 745 750
Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val
755 760 765
Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp
770 775 780
Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys
785 790 795 800
Gln Ser Ile Ile Ser Thr Ser Pro Gln
805
<210> 23
<211> 789
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL7IL12bGMCSF蛋白质异二聚体
<400> 23
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Glu Cys His Ile Lys Asp Lys Glu Gly Lys Ala Tyr Glu Ser Val Leu
210 215 220
Met Ile Ser Ile Asp Glu Leu Asp Lys Met Thr Gly Thr Asp Ser Asn
225 230 235 240
Cys Pro Asn Asn Glu Pro Asn Phe Phe Arg Lys His Val Cys Asp Asp
245 250 255
Thr Lys Glu Ala Ala Phe Leu Asn Arg Ala Ala Arg Lys Leu Lys Gln
260 265 270
Phe Leu Lys Met Asn Ile Ser Glu Glu Phe Asn Val His Leu Leu Thr
275 280 285
Val Ser Gln Gly Thr Gln Thr Leu Val Asn Cys Thr Ser Lys Glu Glu
290 295 300
Lys Asn Val Lys Glu Gln Lys Lys Asn Asp Ala Cys Phe Leu Lys Arg
305 310 315 320
Leu Leu Arg Glu Ile Lys Thr Cys Trp Asn Lys Ile Leu Lys Gly Ser
325 330 335
Ile Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp
340 345 350
Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro
355 360 365
Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile
370 375 380
Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala
385 390 395 400
Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His
405 410 415
Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu
420 425 430
Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr
435 440 445
Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu
450 455 460
Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val
465 470 475 480
Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln
485 490 495
Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys
500 505 510
Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg
515 520 525
Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp
530 535 540
Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys
545 550 555 560
Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr
565 570 575
Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys
580 585 590
Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala
595 600 605
Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn
610 615 620
Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys
625 630 635 640
Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly
645 650 655
Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile
660 665 670
Thr Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu
675 680 685
Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val
690 695 700
Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg
705 710 715 720
Leu Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys
725 730 735
Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro
740 745 750
Thr Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe
755 760 765
Ile Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys
770 775 780
Lys Pro Gly Gln Lys
785
<210> 24
<211> 793
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL15IL12bGMCSF蛋白质异二聚体
<400> 24
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Gly Ile His Val Phe Ile Leu Gly Cys Val Ser Val Gly Leu Pro Lys
210 215 220
Thr Glu Ala Asn Trp Ile Asp Val Arg Tyr Asp Leu Glu Lys Ile Glu
225 230 235 240
Ser Leu Ile Gln Ser Ile His Ile Asp Thr Thr Leu Tyr Thr Asp Ser
245 250 255
Asp Phe His Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu
260 265 270
Glu Leu Gln Val Ile Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu
275 280 285
Thr Val Arg Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn
290 295 300
Lys Asn Val Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu
305 310 315 320
Lys Thr Phe Thr Glu Phe Leu Gln Ser Phe Ile Arg Ile Val Gln Met
325 330 335
Phe Ile Asn Thr Ser Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val
340 345 350
Glu Val Asp Trp Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr
355 360 365
Cys Asp Thr Pro Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg
370 375 380
His Gly Val Ile Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu
385 390 395 400
Phe Leu Asp Ala Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu
405 410 415
Ser His Ser His Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser
420 425 430
Thr Glu Ile Leu Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu
435 440 445
Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg
450 455 460
Asn Met Asp Leu Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp
465 470 475 480
Ser Arg Ala Val Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val
485 490 495
Thr Leu Asp Gln Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu
500 505 510
Asp Val Thr Cys Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala
515 520 525
Leu Glu Ala Arg Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe
530 535 540
Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met
545 550 555 560
Lys Pro Leu Lys Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp
565 570 575
Ser Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg
580 585 590
Ile Gln Arg Lys Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn
595 600 605
Gln Lys Gly Ala Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys
610 615 620
Lys Gly Gly Asn Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser
625 630 635 640
Ser Cys Ser Lys Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly
645 650 655
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr
660 665 670
Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val Glu Ala Ile
675 680 685
Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu
690 695 700
Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys
705 710 715 720
Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe
725 730 735
Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr
740 745 750
Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr
755 760 765
Tyr Ala Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro
770 775 780
Phe Glu Cys Lys Lys Pro Gly Gln Lys
785 790
<210> 25
<211> 789
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL21IL12bGMCSF蛋白质异二聚体
<400> 25
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
His Lys Ser Ser Pro Gln Gly Pro Asp Arg Leu Leu Ile Arg Leu Arg
210 215 220
His Leu Ile Asp Ile Val Glu Gln Leu Lys Ile Tyr Glu Asn Asp Leu
225 230 235 240
Asp Pro Glu Leu Leu Ser Ala Pro Gln Asp Val Lys Gly His Cys Glu
245 250 255
His Ala Ala Phe Ala Cys Phe Gln Lys Ala Lys Leu Lys Pro Ser Asn
260 265 270
Pro Gly Asn Asn Lys Thr Phe Ile Ile Asp Leu Val Ala Gln Leu Arg
275 280 285
Arg Arg Leu Pro Ala Arg Arg Gly Gly Lys Lys Gln Lys His Ile Ala
290 295 300
Lys Cys Pro Ser Cys Asp Ser Tyr Glu Lys Arg Thr Pro Lys Glu Phe
305 310 315 320
Leu Glu Arg Leu Lys Trp Leu Leu Gln Lys Met Ile His Gln His Leu
325 330 335
Ser Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp
340 345 350
Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro
355 360 365
Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile
370 375 380
Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala
385 390 395 400
Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His
405 410 415
Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu
420 425 430
Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr
435 440 445
Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu
450 455 460
Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val
465 470 475 480
Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln
485 490 495
Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys
500 505 510
Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg
515 520 525
Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp
530 535 540
Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys
545 550 555 560
Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr
565 570 575
Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys
580 585 590
Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala
595 600 605
Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn
610 615 620
Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys
625 630 635 640
Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly
645 650 655
Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile
660 665 670
Thr Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu
675 680 685
Asn Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val
690 695 700
Val Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg
705 710 715 720
Leu Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys
725 730 735
Gly Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro
740 745 750
Thr Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe
755 760 765
Ile Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys
770 775 780
Lys Pro Gly Gln Lys
785
<210> 26
<211> 891
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL2IL12bFLT3L蛋白质异二聚体
<400> 26
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
210 215 220
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
225 230 235 240
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
245 250 255
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
260 265 270
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
275 280 285
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
290 295 300
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
305 310 315 320
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
325 330 335
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
340 345 350
Ser Thr Ser Pro Gln Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val
355 360 365
Glu Val Asp Trp Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr
370 375 380
Cys Asp Thr Pro Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg
385 390 395 400
His Gly Val Ile Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu
405 410 415
Phe Leu Asp Ala Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu
420 425 430
Ser His Ser His Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser
435 440 445
Thr Glu Ile Leu Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu
450 455 460
Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg
465 470 475 480
Asn Met Asp Leu Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp
485 490 495
Ser Arg Ala Val Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val
500 505 510
Thr Leu Asp Gln Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu
515 520 525
Asp Val Thr Cys Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala
530 535 540
Leu Glu Ala Arg Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe
545 550 555 560
Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met
565 570 575
Lys Pro Leu Lys Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp
580 585 590
Ser Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg
595 600 605
Ile Gln Arg Lys Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn
610 615 620
Gln Lys Gly Ala Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys
625 630 635 640
Lys Gly Gly Asn Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser
645 650 655
Ser Cys Ser Lys Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly
660 665 670
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro
675 680 685
Asp Cys Tyr Phe Ser His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys
690 695 700
Phe Arg Glu Leu Thr Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val
705 710 715 720
Ala Val Asn Leu Gln Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu
725 730 735
Phe Leu Ala Gln Arg Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser
740 745 750
Lys Met Gln Thr Leu Leu Glu Asp Val Asn Thr Glu Ile His Phe Val
755 760 765
Thr Ser Cys Thr Phe Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln
770 775 780
Thr Asn Ile Ser His Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala
785 790 795 800
Leu Lys Pro Cys Ile Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu
805 810 815
Glu Val Gln Cys Gln Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser
820 825 830
Pro Ile Ala Leu Glu Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gln
835 840 845
Leu Leu Leu Leu Leu Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu
850 855 860
Ala Ala Ala Trp Gly Leu Arg Trp Gln Arg Ala Arg Arg Arg Gly Glu
865 870 875 880
Leu His Pro Gly Val Pro Leu Pro Ser His Pro
885 890
<210> 27
<211> 871
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL7IL12bFLT3L蛋白质异二聚体
<400> 27
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Glu Cys His Ile Lys Asp Lys Glu Gly Lys Ala Tyr Glu Ser Val Leu
210 215 220
Met Ile Ser Ile Asp Glu Leu Asp Lys Met Thr Gly Thr Asp Ser Asn
225 230 235 240
Cys Pro Asn Asn Glu Pro Asn Phe Phe Arg Lys His Val Cys Asp Asp
245 250 255
Thr Lys Glu Ala Ala Phe Leu Asn Arg Ala Ala Arg Lys Leu Lys Gln
260 265 270
Phe Leu Lys Met Asn Ile Ser Glu Glu Phe Asn Val His Leu Leu Thr
275 280 285
Val Ser Gln Gly Thr Gln Thr Leu Val Asn Cys Thr Ser Lys Glu Glu
290 295 300
Lys Asn Val Lys Glu Gln Lys Lys Asn Asp Ala Cys Phe Leu Lys Arg
305 310 315 320
Leu Leu Arg Glu Ile Lys Thr Cys Trp Asn Lys Ile Leu Lys Gly Ser
325 330 335
Ile Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp
340 345 350
Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro
355 360 365
Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile
370 375 380
Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala
385 390 395 400
Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His
405 410 415
Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu
420 425 430
Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr
435 440 445
Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu
450 455 460
Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val
465 470 475 480
Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln
485 490 495
Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys
500 505 510
Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg
515 520 525
Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp
530 535 540
Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys
545 550 555 560
Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr
565 570 575
Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys
580 585 590
Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala
595 600 605
Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn
610 615 620
Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys
625 630 635 640
Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly
645 650 655
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro Asp Cys Tyr Phe
660 665 670
Ser His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys Phe Arg Glu Leu
675 680 685
Thr Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val Ala Val Asn Leu
690 695 700
Gln Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu Phe Leu Ala Gln
705 710 715 720
Arg Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser Lys Met Gln Thr
725 730 735
Leu Leu Glu Asp Val Asn Thr Glu Ile His Phe Val Thr Ser Cys Thr
740 745 750
Phe Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln Thr Asn Ile Ser
755 760 765
His Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala Leu Lys Pro Cys
770 775 780
Ile Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu Glu Val Gln Cys
785 790 795 800
Gln Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser Pro Ile Ala Leu
805 810 815
Glu Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gln Leu Leu Leu Leu
820 825 830
Leu Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu Ala Ala Ala Trp
835 840 845
Gly Leu Arg Trp Gln Arg Ala Arg Arg Arg Gly Glu Leu His Pro Gly
850 855 860
Val Pro Leu Pro Ser His Pro
865 870
<210> 28
<211> 875
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL15IL12bFLT3L蛋白质异二聚体
<400> 28
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Gly Ile His Val Phe Ile Leu Gly Cys Val Ser Val Gly Leu Pro Lys
210 215 220
Thr Glu Ala Asn Trp Ile Asp Val Arg Tyr Asp Leu Glu Lys Ile Glu
225 230 235 240
Ser Leu Ile Gln Ser Ile His Ile Asp Thr Thr Leu Tyr Thr Asp Ser
245 250 255
Asp Phe His Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu
260 265 270
Glu Leu Gln Val Ile Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu
275 280 285
Thr Val Arg Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn
290 295 300
Lys Asn Val Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu
305 310 315 320
Lys Thr Phe Thr Glu Phe Leu Gln Ser Phe Ile Arg Ile Val Gln Met
325 330 335
Phe Ile Asn Thr Ser Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val
340 345 350
Glu Val Asp Trp Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr
355 360 365
Cys Asp Thr Pro Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg
370 375 380
His Gly Val Ile Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu
385 390 395 400
Phe Leu Asp Ala Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu
405 410 415
Ser His Ser His Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser
420 425 430
Thr Glu Ile Leu Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu
435 440 445
Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg
450 455 460
Asn Met Asp Leu Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp
465 470 475 480
Ser Arg Ala Val Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val
485 490 495
Thr Leu Asp Gln Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu
500 505 510
Asp Val Thr Cys Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala
515 520 525
Leu Glu Ala Arg Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe
530 535 540
Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met
545 550 555 560
Lys Pro Leu Lys Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp
565 570 575
Ser Trp Ser Thr Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg
580 585 590
Ile Gln Arg Lys Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn
595 600 605
Gln Lys Gly Ala Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys
610 615 620
Lys Gly Gly Asn Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser
625 630 635 640
Ser Cys Ser Lys Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly
645 650 655
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro
660 665 670
Asp Cys Tyr Phe Ser His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys
675 680 685
Phe Arg Glu Leu Thr Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val
690 695 700
Ala Val Asn Leu Gln Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu
705 710 715 720
Phe Leu Ala Gln Arg Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser
725 730 735
Lys Met Gln Thr Leu Leu Glu Asp Val Asn Thr Glu Ile His Phe Val
740 745 750
Thr Ser Cys Thr Phe Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln
755 760 765
Thr Asn Ile Ser His Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala
770 775 780
Leu Lys Pro Cys Ile Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu
785 790 795 800
Glu Val Gln Cys Gln Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser
805 810 815
Pro Ile Ala Leu Glu Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gln
820 825 830
Leu Leu Leu Leu Leu Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu
835 840 845
Ala Ala Ala Trp Gly Leu Arg Trp Gln Arg Ala Arg Arg Arg Gly Glu
850 855 860
Leu His Pro Gly Val Pro Leu Pro Ser His Pro
865 870 875
<210> 29
<211> 871
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12aIL21IL12bFLT3L蛋白质异二聚体
<400> 29
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
His Lys Ser Ser Pro Gln Gly Pro Asp Arg Leu Leu Ile Arg Leu Arg
210 215 220
His Leu Ile Asp Ile Val Glu Gln Leu Lys Ile Tyr Glu Asn Asp Leu
225 230 235 240
Asp Pro Glu Leu Leu Ser Ala Pro Gln Asp Val Lys Gly His Cys Glu
245 250 255
His Ala Ala Phe Ala Cys Phe Gln Lys Ala Lys Leu Lys Pro Ser Asn
260 265 270
Pro Gly Asn Asn Lys Thr Phe Ile Ile Asp Leu Val Ala Gln Leu Arg
275 280 285
Arg Arg Leu Pro Ala Arg Arg Gly Gly Lys Lys Gln Lys His Ile Ala
290 295 300
Lys Cys Pro Ser Cys Asp Ser Tyr Glu Lys Arg Thr Pro Lys Glu Phe
305 310 315 320
Leu Glu Arg Leu Lys Trp Leu Leu Gln Lys Met Ile His Gln His Leu
325 330 335
Ser Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp
340 345 350
Thr Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro
355 360 365
Glu Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile
370 375 380
Gly Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala
385 390 395 400
Gly Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His
405 410 415
Leu Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu
420 425 430
Lys Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr
435 440 445
Ser Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu
450 455 460
Lys Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val
465 470 475 480
Thr Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln
485 490 495
Arg Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys
500 505 510
Pro Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg
515 520 525
Gln Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp
530 535 540
Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys
545 550 555 560
Asn Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr
565 570 575
Pro His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys
580 585 590
Lys Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala
595 600 605
Phe Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn
610 615 620
Val Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys
625 630 635 640
Trp Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly
645 650 655
Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro Asp Cys Tyr Phe
660 665 670
Ser His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys Phe Arg Glu Leu
675 680 685
Thr Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val Ala Val Asn Leu
690 695 700
Gln Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu Phe Leu Ala Gln
705 710 715 720
Arg Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser Lys Met Gln Thr
725 730 735
Leu Leu Glu Asp Val Asn Thr Glu Ile His Phe Val Thr Ser Cys Thr
740 745 750
Phe Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln Thr Asn Ile Ser
755 760 765
His Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala Leu Lys Pro Cys
770 775 780
Ile Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu Glu Val Gln Cys
785 790 795 800
Gln Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser Pro Ile Ala Leu
805 810 815
Glu Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gln Leu Leu Leu Leu
820 825 830
Leu Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu Ala Ala Ala Trp
835 840 845
Gly Leu Arg Trp Gln Arg Ala Arg Arg Arg Gly Glu Leu His Pro Gly
850 855 860
Val Pro Leu Pro Ser His Pro
865 870
<210> 30
<211> 793
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL2IL12bGMCSF蛋白质异二聚体
<400> 30
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu
210 215 220
Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile
225 230 235 240
Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe
245 250 255
Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu
260 265 270
Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys
275 280 285
Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile
290 295 300
Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala
305 310 315 320
Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe
325 330 335
Cys Gln Ser Ile Ile Ser Thr Leu Thr Ile Trp Glu Leu Lys Lys Asp
340 345 350
Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met
355 360 365
Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile Thr Trp Thr
370 375 380
Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr Ile
385 390 395 400
Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys His Lys Gly
405 410 415
Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp
420 425 430
Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu Pro Lys Asn
435 440 445
Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr
450 455 460
Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe Ser Val Lys
465 470 475 480
Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys Gly Ala Ala
485 490 495
Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr
500 505 510
Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser
515 520 525
Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu
530 535 540
Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro
545 550 555 560
Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg Gln Val Glu
565 570 575
Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe
580 585 590
Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys Arg Glu Lys
595 600 605
Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val Ile Cys Arg
610 615 620
Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr Tyr Ser Ser
625 630 635 640
Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly Gly Gly Ser
645 650 655
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala Arg Ser Pro
660 665 670
Ser Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile Gln Glu Ala
675 680 685
Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met Asn Glu
690 695 700
Thr Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu Pro Thr Cys
705 710 715 720
Leu Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg Gly Ser Leu
725 730 735
Thr Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr Lys Gln
740 745 750
His Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln Ile Ile Thr
755 760 765
Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val Ile Pro
770 775 780
Phe Asp Cys Trp Glu Pro Val Gln Glu
785 790
<210> 31
<211> 812
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL7IL12bGMCSF蛋白质异二聚体
<400> 31
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr
210 215 220
Glu Ser Val Leu Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys
225 230 235 240
Glu Ile Gly Ser Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg
245 250 255
His Ile Cys Asp Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala
260 265 270
Arg Lys Leu Arg Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp
275 280 285
Leu His Leu Leu Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys
290 295 300
Thr Gly Gln Val Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln
305 310 315 320
Pro Thr Lys Ser Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys
325 330 335
Leu Asn Asp Leu Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr
340 345 350
Cys Trp Asn Lys Ile Leu Met Gly Thr Lys Glu His Ile Trp Glu Leu
355 360 365
Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro
370 375 380
Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile
385 390 395 400
Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr
405 410 415
Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys
420 425 430
His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys
435 440 445
Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu
450 455 460
Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly
465 470 475 480
Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe
485 490 495
Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys
500 505 510
Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu
515 520 525
Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala
530 535 540
Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu
545 550 555 560
Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys
565 570 575
Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg
580 585 590
Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His
595 600 605
Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys
610 615 620
Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val
625 630 635 640
Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr
645 650 655
Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly
660 665 670
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala
675 680 685
Arg Ser Pro Ser Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile
690 695 700
Gln Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu
705 710 715 720
Met Asn Glu Thr Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu
725 730 735
Pro Thr Cys Leu Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg
740 745 750
Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His
755 760 765
Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln
770 775 780
Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu
785 790 795 800
Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu
805 810
<210> 32
<211> 793
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL15IL12bGMCSF蛋白质异二聚体
<400> 32
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Gly Ile His Val Phe Ile Leu Gly Cys Phe Ser Ala
210 215 220
Gly Leu Pro Lys Thr Glu Ala Asn Trp Val Asn Val Ile Ser Asp Leu
225 230 235 240
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
245 250 255
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
260 265 270
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
275 280 285
Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
290 295 300
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
305 310 315 320
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
325 330 335
Ile Val Gln Met Phe Ile Asn Thr Ser Ile Trp Glu Leu Lys Lys Asp
340 345 350
Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met
355 360 365
Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile Thr Trp Thr
370 375 380
Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr Ile
385 390 395 400
Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys His Lys Gly
405 410 415
Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp
420 425 430
Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu Pro Lys Asn
435 440 445
Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr
450 455 460
Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe Ser Val Lys
465 470 475 480
Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys Gly Ala Ala
485 490 495
Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr
500 505 510
Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser
515 520 525
Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu
530 535 540
Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro
545 550 555 560
Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg Gln Val Glu
565 570 575
Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe
580 585 590
Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys Arg Glu Lys
595 600 605
Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val Ile Cys Arg
610 615 620
Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr Tyr Ser Ser
625 630 635 640
Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly Gly Gly Ser
645 650 655
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala Arg Ser Pro
660 665 670
Ser Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile Gln Glu Ala
675 680 685
Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met Asn Glu
690 695 700
Thr Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu Pro Thr Cys
705 710 715 720
Leu Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg Gly Ser Leu
725 730 735
Thr Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr Lys Gln
740 745 750
His Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln Ile Ile Thr
755 760 765
Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val Ile Pro
770 775 780
Phe Asp Cys Trp Glu Pro Val Gln Glu
785 790
<210> 33
<211> 793
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL21IL12bGMCSF蛋白质异二聚体
<400> 33
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Gln Gly Gln Asp Arg His Met Ile Arg Met Arg Gln
210 215 220
Leu Ile Asp Ile Val Asp Gln Leu Lys Asn Tyr Val Asn Asp Leu Val
225 230 235 240
Pro Glu Phe Leu Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu Trp
245 250 255
Ser Ala Phe Ser Cys Phe Gln Lys Ala Gln Leu Lys Ser Ala Asn Thr
260 265 270
Gly Asn Asn Glu Arg Ile Ile Asn Val Ser Ile Lys Lys Leu Lys Arg
275 280 285
Lys Pro Pro Ser Thr Asn Ala Gly Arg Arg Gln Lys His Arg Leu Thr
290 295 300
Cys Pro Ser Cys Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe Leu
305 310 315 320
Glu Arg Phe Lys Ser Leu Leu Gln Lys Met Ile His Gln His Leu Ser
325 330 335
Ser Arg Thr His Gly Ser Glu Asp Ser Ile Trp Glu Leu Lys Lys Asp
340 345 350
Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met
355 360 365
Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile Thr Trp Thr
370 375 380
Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr Ile
385 390 395 400
Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys His Lys Gly
405 410 415
Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp
420 425 430
Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu Pro Lys Asn
435 440 445
Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr
450 455 460
Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe Ser Val Lys
465 470 475 480
Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys Gly Ala Ala
485 490 495
Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr
500 505 510
Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser
515 520 525
Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu
530 535 540
Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro
545 550 555 560
Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg Gln Val Glu
565 570 575
Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe
580 585 590
Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys Arg Glu Lys
595 600 605
Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val Ile Cys Arg
610 615 620
Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr Tyr Ser Ser
625 630 635 640
Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly Gly Gly Ser
645 650 655
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Ala Arg Ser Pro
660 665 670
Ser Pro Ser Thr Gln Pro Trp Glu His Val Asn Ala Ile Gln Glu Ala
675 680 685
Arg Arg Leu Leu Asn Leu Ser Arg Asp Thr Ala Ala Glu Met Asn Glu
690 695 700
Thr Val Glu Val Ile Ser Glu Met Phe Asp Leu Gln Glu Pro Thr Cys
705 710 715 720
Leu Gln Thr Arg Leu Glu Leu Tyr Lys Gln Gly Leu Arg Gly Ser Leu
725 730 735
Thr Lys Leu Lys Gly Pro Leu Thr Met Met Ala Ser His Tyr Lys Gln
740 745 750
His Cys Pro Pro Thr Pro Glu Thr Ser Cys Ala Thr Gln Ile Ile Thr
755 760 765
Phe Glu Ser Phe Lys Glu Asn Leu Lys Asp Phe Leu Leu Val Ile Pro
770 775 780
Phe Asp Cys Trp Glu Pro Val Gln Glu
785 790
<210> 34
<211> 875
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL2IL12bFLT3L蛋白质异二聚体
<400> 34
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu
210 215 220
Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile
225 230 235 240
Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe
245 250 255
Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu
260 265 270
Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys
275 280 285
Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile
290 295 300
Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala
305 310 315 320
Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe
325 330 335
Cys Gln Ser Ile Ile Ser Thr Leu Thr Ile Trp Glu Leu Lys Lys Asp
340 345 350
Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met
355 360 365
Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile Thr Trp Thr
370 375 380
Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr Ile
385 390 395 400
Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys His Lys Gly
405 410 415
Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp
420 425 430
Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu Pro Lys Asn
435 440 445
Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr
450 455 460
Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe Ser Val Lys
465 470 475 480
Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys Gly Ala Ala
485 490 495
Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr
500 505 510
Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser
515 520 525
Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu
530 535 540
Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro
545 550 555 560
Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg Gln Val Glu
565 570 575
Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe
580 585 590
Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys Arg Glu Lys
595 600 605
Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val Ile Cys Arg
610 615 620
Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr Tyr Ser Ser
625 630 635 640
Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly Gly Gly Ser
645 650 655
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gln Asp Cys Ser Phe
660 665 670
Gln His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu
675 680 685
Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu
690 695 700
Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln
705 710 715 720
Arg Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly
725 730 735
Leu Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala
740 745 750
Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser
755 760 765
Arg Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp
770 775 780
Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro
785 790 795 800
Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala
805 810 815
Thr Ala Pro Thr Ala Pro Gln Pro Pro Leu Leu Leu Leu Leu Leu Leu
820 825 830
Pro Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu His Trp Gln
835 840 845
Arg Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gln Val Pro Pro Val
850 855 860
Pro Ser Pro Gln Asp Leu Leu Leu Val Glu His
865 870 875
<210> 35
<211> 894
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL7IL12bFLT3L蛋白质异二聚体
<400> 35
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr
210 215 220
Glu Ser Val Leu Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys
225 230 235 240
Glu Ile Gly Ser Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg
245 250 255
His Ile Cys Asp Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala
260 265 270
Arg Lys Leu Arg Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp
275 280 285
Leu His Leu Leu Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys
290 295 300
Thr Gly Gln Val Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln
305 310 315 320
Pro Thr Lys Ser Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys
325 330 335
Leu Asn Asp Leu Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr
340 345 350
Cys Trp Asn Lys Ile Leu Met Gly Thr Lys Glu His Ile Trp Glu Leu
355 360 365
Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro
370 375 380
Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile
385 390 395 400
Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr
405 410 415
Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys
420 425 430
His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys
435 440 445
Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu
450 455 460
Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly
465 470 475 480
Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe
485 490 495
Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys
500 505 510
Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu
515 520 525
Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala
530 535 540
Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu
545 550 555 560
Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys
565 570 575
Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg
580 585 590
Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His
595 600 605
Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys
610 615 620
Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val
625 630 635 640
Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr
645 650 655
Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly
660 665 670
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gln Asp
675 680 685
Cys Ser Phe Gln His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile
690 695 700
Arg Glu Leu Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala
705 710 715 720
Ser Asn Leu Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val
725 730 735
Leu Ala Gln Arg Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys
740 745 750
Met Gln Gly Leu Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr
755 760 765
Lys Cys Ala Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr
770 775 780
Asn Ile Ser Arg Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu
785 790 795 800
Lys Pro Trp Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln
805 810 815
Cys Gln Pro Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro
820 825 830
Leu Glu Ala Thr Ala Pro Thr Ala Pro Gln Pro Pro Leu Leu Leu Leu
835 840 845
Leu Leu Leu Pro Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu
850 855 860
His Trp Gln Arg Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gln Val
865 870 875 880
Pro Pro Val Pro Ser Pro Gln Asp Leu Leu Leu Val Glu His
885 890
<210> 36
<211> 875
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL15IL12bFLT3L蛋白质异二聚体
<400> 36
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Gly Ile His Val Phe Ile Leu Gly Cys Phe Ser Ala
210 215 220
Gly Leu Pro Lys Thr Glu Ala Asn Trp Val Asn Val Ile Ser Asp Leu
225 230 235 240
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
245 250 255
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
260 265 270
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
275 280 285
Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
290 295 300
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
305 310 315 320
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
325 330 335
Ile Val Gln Met Phe Ile Asn Thr Ser Ile Trp Glu Leu Lys Lys Asp
340 345 350
Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met
355 360 365
Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile Thr Trp Thr
370 375 380
Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr Ile
385 390 395 400
Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys His Lys Gly
405 410 415
Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp
420 425 430
Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu Pro Lys Asn
435 440 445
Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr
450 455 460
Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe Ser Val Lys
465 470 475 480
Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys Gly Ala Ala
485 490 495
Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr
500 505 510
Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser
515 520 525
Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu
530 535 540
Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro
545 550 555 560
Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg Gln Val Glu
565 570 575
Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe
580 585 590
Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys Arg Glu Lys
595 600 605
Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val Ile Cys Arg
610 615 620
Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr Tyr Ser Ser
625 630 635 640
Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly Gly Gly Ser
645 650 655
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gln Asp Cys Ser Phe
660 665 670
Gln His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu
675 680 685
Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu
690 695 700
Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln
705 710 715 720
Arg Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly
725 730 735
Leu Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala
740 745 750
Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser
755 760 765
Arg Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp
770 775 780
Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro
785 790 795 800
Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala
805 810 815
Thr Ala Pro Thr Ala Pro Gln Pro Pro Leu Leu Leu Leu Leu Leu Leu
820 825 830
Pro Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu His Trp Gln
835 840 845
Arg Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gln Val Pro Pro Val
850 855 860
Pro Ser Pro Gln Asp Leu Leu Leu Val Glu His
865 870 875
<210> 37
<211> 875
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12aIL21IL12bFLT3L蛋白质异二聚体
<400> 37
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Gln Gly Gln Asp Arg His Met Ile Arg Met Arg Gln
210 215 220
Leu Ile Asp Ile Val Asp Gln Leu Lys Asn Tyr Val Asn Asp Leu Val
225 230 235 240
Pro Glu Phe Leu Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu Trp
245 250 255
Ser Ala Phe Ser Cys Phe Gln Lys Ala Gln Leu Lys Ser Ala Asn Thr
260 265 270
Gly Asn Asn Glu Arg Ile Ile Asn Val Ser Ile Lys Lys Leu Lys Arg
275 280 285
Lys Pro Pro Ser Thr Asn Ala Gly Arg Arg Gln Lys His Arg Leu Thr
290 295 300
Cys Pro Ser Cys Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe Leu
305 310 315 320
Glu Arg Phe Lys Ser Leu Leu Gln Lys Met Ile His Gln His Leu Ser
325 330 335
Ser Arg Thr His Gly Ser Glu Asp Ser Ile Trp Glu Leu Lys Lys Asp
340 345 350
Val Tyr Val Val Glu Leu Asp Trp Tyr Pro Asp Ala Pro Gly Glu Met
355 360 365
Val Val Leu Thr Cys Asp Thr Pro Glu Glu Asp Gly Ile Thr Trp Thr
370 375 380
Leu Asp Gln Ser Ser Glu Val Leu Gly Ser Gly Lys Thr Leu Thr Ile
385 390 395 400
Gln Val Lys Glu Phe Gly Asp Ala Gly Gln Tyr Thr Cys His Lys Gly
405 410 415
Gly Glu Val Leu Ser His Ser Leu Leu Leu Leu His Lys Lys Glu Asp
420 425 430
Gly Ile Trp Ser Thr Asp Ile Leu Lys Asp Gln Lys Glu Pro Lys Asn
435 440 445
Lys Thr Phe Leu Arg Cys Glu Ala Lys Asn Tyr Ser Gly Arg Phe Thr
450 455 460
Cys Trp Trp Leu Thr Thr Ile Ser Thr Asp Leu Thr Phe Ser Val Lys
465 470 475 480
Ser Ser Arg Gly Ser Ser Asp Pro Gln Gly Val Thr Cys Gly Ala Ala
485 490 495
Thr Leu Ser Ala Glu Arg Val Arg Gly Asp Asn Lys Glu Tyr Glu Tyr
500 505 510
Ser Val Glu Cys Gln Glu Asp Ser Ala Cys Pro Ala Ala Glu Glu Ser
515 520 525
Leu Pro Ile Glu Val Met Val Asp Ala Val His Lys Leu Lys Tyr Glu
530 535 540
Asn Tyr Thr Ser Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro
545 550 555 560
Pro Lys Asn Leu Gln Leu Lys Pro Leu Lys Asn Ser Arg Gln Val Glu
565 570 575
Val Ser Trp Glu Tyr Pro Asp Thr Trp Ser Thr Pro His Ser Tyr Phe
580 585 590
Ser Leu Thr Phe Cys Val Gln Val Gln Gly Lys Ser Lys Arg Glu Lys
595 600 605
Lys Asp Arg Val Phe Thr Asp Lys Thr Ser Ala Thr Val Ile Cys Arg
610 615 620
Lys Asn Ala Ser Ile Ser Val Arg Ala Gln Asp Arg Tyr Tyr Ser Ser
625 630 635 640
Ser Trp Ser Glu Trp Ala Ser Val Pro Cys Ser Gly Gly Gly Gly Ser
645 650 655
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Thr Gln Asp Cys Ser Phe
660 665 670
Gln His Ser Pro Ile Ser Ser Asp Phe Ala Val Lys Ile Arg Glu Leu
675 680 685
Ser Asp Tyr Leu Leu Gln Asp Tyr Pro Val Thr Val Ala Ser Asn Leu
690 695 700
Gln Asp Glu Glu Leu Cys Gly Gly Leu Trp Arg Leu Val Leu Ala Gln
705 710 715 720
Arg Trp Met Glu Arg Leu Lys Thr Val Ala Gly Ser Lys Met Gln Gly
725 730 735
Leu Leu Glu Arg Val Asn Thr Glu Ile His Phe Val Thr Lys Cys Ala
740 745 750
Phe Gln Pro Pro Pro Ser Cys Leu Arg Phe Val Gln Thr Asn Ile Ser
755 760 765
Arg Leu Leu Gln Glu Thr Ser Glu Gln Leu Val Ala Leu Lys Pro Trp
770 775 780
Ile Thr Arg Gln Asn Phe Ser Arg Cys Leu Glu Leu Gln Cys Gln Pro
785 790 795 800
Asp Ser Ser Thr Leu Pro Pro Pro Trp Ser Pro Arg Pro Leu Glu Ala
805 810 815
Thr Ala Pro Thr Ala Pro Gln Pro Pro Leu Leu Leu Leu Leu Leu Leu
820 825 830
Pro Val Gly Leu Leu Leu Leu Ala Ala Ala Trp Cys Leu His Trp Gln
835 840 845
Arg Thr Arg Arg Arg Thr Pro Arg Pro Gly Glu Gln Val Pro Pro Val
850 855 860
Pro Ser Pro Gln Asp Leu Leu Leu Val Glu His
865 870 875
<210> 38
<211> 2616
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL2IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 38
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc gcacccactt caagctccac ttcaagctct 720
acagcggaag cacagcagca gcagcagcag cagcagcagc agcagcagca cctggagcag 780
ctgttgatgg acctacagga gctcctgagc aggatggaga attacaggaa cctgaaactc 840
cccaggatgc tcaccttcaa attttacttg cccaagcagg ccacagaatt gaaagatctt 900
cagtgcctag aagatgaact tggacctctg cggcatgttc tggatttgac tcaaagcaaa 960
agctttcaat tggaagatgc tgagaatttc atcagcaata tcagagtaac tgttgtaaaa 1020
ctaaagggct ctgacaacac atttgagtgc caattcgatg atgagtcagc aactgtggtg 1080
gactttctga ggagatggat agccttctgt caaagcatca tctcaacaag ccctcaagag 1140
ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtgtcct 1200
cagaagctaa ccatctcctg gtttgccatc gttttgctgg tgtctccact catggccatg 1260
tgggagctgg agaaagacgt ttatgttgta gaggtggact ggactcccga tgcccctgga 1320
gaaacagtga acctcacctg tgacacgcct gaagaagatg acatcacctg gacctcagac 1380
cagagacatg gagtcatagg ctctggaaag accctgacca tcactgtcaa agagtttcta 1440
gatgctggcc agtacacctg ccacaaagga ggcgagactc tgagccactc acatctgctg 1500
ctccacaaga aggaaaatgg aatttggtcc actgaaattt taaaaaattt caaaaacaag 1560
actttcctga agtgtgaagc accaaattac tccggacggt tcacgtgctc atggctggtg 1620
caaagaaaca tggacttgaa gttcaacatc aagagcagta gcagttcccc tgactctcgg 1680
gcagtgacat gtggaatggc gtctctgtct gcagagaagg tcacactgga ccaaagggac 1740
tatgagaagt attcagtgtc ctgccaggag gatgtcacct gcccaactgc cgaggagacc 1800
ctgcccattg aactggcgtt ggaagcacgg cagcagaata aatatgagaa ctacagcacc 1860
agcttcttca tcagggacat catcaaacca gacccgccca agaacttgca gatgaagcct 1920
ttgaagaact cacaggtgga ggtcagctgg gagtaccctg actcctggag cactccccat 1980
tcctacttct ccctcaagtt ctttgttcga atccagcgca agaaagaaaa gatgaaggag 2040
acagaggagg ggtgtaacca gaaaggtgcg ttcctcgtag agaagacatc taccgaagtc 2100
caatgcaaag gcgggaatgt ctgcgtgcaa gctcaggatc gctattacaa ttcctcgtgc 2160
agcaagtggg catgtgttcc ctgcagggtc cgatccggag gcggtggaag tggcggtgga 2220
ggctctggag gtggcggaag cgcacccacc cgctcaccca tcactgtcac ccggccttgg 2280
aagcatgtag aggccatcaa agaagccctg aacctcctgg atgacatgcc tgtcacgttg 2340
aatgaagagg tagaagtcgt ctctaacgag ttctccttca agaagctaac atgtgtgcag 2400
acccgcctga agatattcga gcagggtcta cggggcaatt tcaccaaact caagggcgcc 2460
ttgaacatga cagccagcta ctaccagaca tactgccccc caactccgga aacggactgt 2520
gaaacacaag ttaccaccta tgcggatttc atagacagcc ttaaaacctt tctgactgat 2580
atcccctttg aatgcaaaaa accaggccaa aaatga 2616
<210> 39
<211> 2610
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL2IL12aIL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 39
atgtacagca tgcagctcgc atcctgtgtc acattgacac ttgtgctcct tgtcaacagc 60
gcacccactt caagctccac ttcaagctct acagcggaag cacagcagca gcagcagcag 120
cagcagcagc agcagcagca cctggagcag ctgttgatgg acctacagga gctcctgagc 180
aggatggaga attacaggaa cctgaaactc cccaggatgc tcaccttcaa attttacttg 240
cccaagcagg ccacagaatt gaaagatctt cagtgcctag aagatgaact tggacctctg 300
cggcatgttc tggatttgac tcaaagcaaa agctttcaat tggaagatgc tgagaatttc 360
atcagcaata tcagagtaac tgttgtaaaa ctaaagggct ctgacaacac atttgagtgc 420
caattcgatg atgagtcagc aactgtggtg gactttctga ggagatggat agccttctgt 480
caaagcatca tctcaacaag ccctcaaggt ggaggaggtt ctggaggcgg tggaagtggt 540
ggcggaggta gcagggtcat tccagtctct ggacctgcca ggtgtcttag ccagtcccga 600
aacctgctga agaccacaga tgacatggtg aagacggcca gagaaaaact gaaacattat 660
tcctgcactg ctgaagacat cgatcatgaa gacatcacac gggaccaaac cagcacattg 720
aagacctgtt taccactgga actacacaag aacgagagtt gcctggctac tagagagact 780
tcttccacaa caagagggag ctgcctgccc ccacagaaga cgtctttgat gatgaccctg 840
tgccttggta gcatctatga ggacttgaag atgtaccaga cagagttcca ggccatcaac 900
gcagcacttc agaatcacaa ccatcagcag atcattctag acaagggcat gctggtggcc 960
atcgatgagc tgatgcagtc tctgaatcat aatggcgaga ctctgcgcca gaaacctcct 1020
gtgggagaag cagaccctta cagagtgaaa atgaagctct gcatcctgct tcacgccttc 1080
agcacccgcg tcgtgaccat caacagggtg atgggctatc tgagctccgc cgagggcaga 1140
ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgtg tcctcagaag 1200
ctaaccatct cctggtttgc catcgttttg ctggtgtctc cactcatggc catgtgggag 1260
ctggagaaag acgtttatgt tgtagaggtg gactggactc ccgatgcccc tggagaaaca 1320
gtgaacctca cctgtgacac gcctgaagaa gatgacatca cctggacctc agaccagaga 1380
catggagtca taggctctgg aaagaccctg accatcactg tcaaagagtt tctagatgct 1440
ggccagtaca cctgccacaa aggaggcgag actctgagcc actcacatct gctgctccac 1500
aagaaggaaa atggaatttg gtccactgaa attttaaaaa atttcaaaaa caagactttc 1560
ctgaagtgtg aagcaccaaa ttactccgga cggttcacgt gctcatggct ggtgcaaaga 1620
aacatggact tgaagttcaa catcaagagc agtagcagtt cccctgactc tcgggcagtg 1680
acatgtggaa tggcgtctct gtctgcagag aaggtcacac tggaccaaag ggactatgag 1740
aagtattcag tgtcctgcca ggaggatgtc acctgcccaa ctgccgagga gaccctgccc 1800
attgaactgg cgttggaagc acggcagcag aataaatatg agaactacag caccagcttc 1860
ttcatcaggg acatcatcaa accagacccg cccaagaact tgcagatgaa gcctttgaag 1920
aactcacagg tggaggtcag ctgggagtac cctgactcct ggagcactcc ccattcctac 1980
ttctccctca agttctttgt tcgaatccag cgcaagaaag aaaagatgaa ggagacagag 2040
gaggggtgta accagaaagg tgcgttcctc gtagagaaga catctaccga agtccaatgc 2100
aaaggcggga atgtctgcgt gcaagctcag gatcgctatt acaattcctc gtgcagcaag 2160
tgggcatgtg ttccctgcag ggtccgatcc ggaggcggtg gaagtggcgg tggaggctct 2220
ggaggtggcg gaagcgcacc cacccgctca cccatcactg tcacccggcc ttggaagcat 2280
gtagaggcca tcaaagaagc cctgaacctc ctggatgaca tgcctgtcac gttgaatgaa 2340
gaggtagaag tcgtctctaa cgagttctcc ttcaagaagc taacatgtgt gcagacccgc 2400
ctgaagatat tcgagcaggg tctacggggc aatttcacca aactcaaggg cgccttgaac 2460
atgacagcca gctactacca gacatactgc cccccaactc cggaaacgga ctgtgaaaca 2520
caagttacca cctatgcgga tttcatagac agccttaaaa cctttctgac tgatatcccc 2580
tttgaatgca aaaaaccagg ccaaaaatga 2610
<210> 40
<211> 2601
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL2GMCSFIL12b蛋白质异二聚体的核苷酸序列
<400> 40
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc gcacccactt caagctccac ttcaagctct 720
acagcggaag cacagcagca gcagcagcag cagcagcagc agcagcagca cctggagcag 780
ctgttgatgg acctacagga gctcctgagc aggatggaga attacaggaa cctgaaactc 840
cccaggatgc tcaccttcaa attttacttg cccaagcagg ccacagaatt gaaagatctt 900
cagtgcctag aagatgaact tggacctctg cggcatgttc tggatttgac tcaaagcaaa 960
agctttcaat tggaagatgc tgagaatttc atcagcaata tcagagtaac tgttgtaaaa 1020
ctaaagggct ctgacaacac atttgagtgc caattcgatg atgagtcagc aactgtggtg 1080
gactttctga ggagatggat agccttctgt caaagcatca tctcaacaag ccctcaagag 1140
ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtggctg 1200
cagaatttac ttttcctggg cattgtggtc tacagcctct cagcacccac ccgctcaccc 1260
atcactgtca cccggccttg gaagcatgta gaggccatca aagaagccct gaacctcctg 1320
gatgacatgc ctgtcacgtt gaatgaagag gtagaagtcg tctctaacga gttctccttc 1380
aagaagctaa catgtgtgca gacccgcctg aagatattcg agcagggtct acggggcaat 1440
ttcaccaaac tcaagggcgc cttgaacatg acagccagct actaccagac atactgcccc 1500
ccaactccgg aaacggactg tgaaacacaa gttaccacct atgcggattt catagacagc 1560
cttaaaacct ttctgactga tatccccttt gaatgcaaaa aaccaggcca aaaaggaggc 1620
ggtggaagtg gcggtggagg ctctggaggt ggcggaagca tgtgggagct ggagaaagac 1680
gtttatgttg tagaggtgga ctggactccc gatgcccctg gagaaacagt gaacctcacc 1740
tgtgacacgc ctgaagaaga tgacatcacc tggacctcag accagagaca tggagtcata 1800
ggctctggaa agaccctgac catcactgtc aaagagtttc tagatgctgg ccagtacacc 1860
tgccacaaag gaggcgagac tctgagccac tcacatctgc tgctccacaa gaaggaaaat 1920
ggaatttggt ccactgaaat tttaaaaaat ttcaaaaaca agactttcct gaagtgtgaa 1980
gcaccaaatt actccggacg gttcacgtgc tcatggctgg tgcaaagaaa catggacttg 2040
aagttcaaca tcaagagcag tagcagttcc cctgactctc gggcagtgac atgtggaatg 2100
gcgtctctgt ctgcagagaa ggtcacactg gaccaaaggg actatgagaa gtattcagtg 2160
tcctgccagg aggatgtcac ctgcccaact gccgaggaga ccctgcccat tgaactggcg 2220
ttggaagcac ggcagcagaa taaatatgag aactacagca ccagcttctt catcagggac 2280
atcatcaaac cagacccgcc caagaacttg cagatgaagc ctttgaagaa ctcacaggtg 2340
gaggtcagct gggagtaccc tgactcctgg agcactcccc attcctactt ctccctcaag 2400
ttctttgttc gaatccagcg caagaaagaa aagatgaagg agacagagga ggggtgtaac 2460
cagaaaggtg cgttcctcgt agagaagaca tctaccgaag tccaatgcaa aggcgggaat 2520
gtctgcgtgc aagctcagga tcgctattac aattcctcgt gcagcaagtg ggcatgtgtt 2580
ccctgcaggg tccgatccta g 2601
<210> 41
<211> 2595
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL2IL12aGMCSFIL12b蛋白质异二聚体的核苷酸序列
<400> 41
atgtacagca tgcagctcgc atcctgtgtc acattgacac ttgtgctcct tgtcaacagc 60
gcacccactt caagctccac ttcaagctct acagcggaag cacagcagca gcagcagcag 120
cagcagcagc agcagcagca cctggagcag ctgttgatgg acctacagga gctcctgagc 180
aggatggaga attacaggaa cctgaaactc cccaggatgc tcaccttcaa attttacttg 240
cccaagcagg ccacagaatt gaaagatctt cagtgcctag aagatgaact tggacctctg 300
cggcatgttc tggatttgac tcaaagcaaa agctttcaat tggaagatgc tgagaatttc 360
atcagcaata tcagagtaac tgttgtaaaa ctaaagggct ctgacaacac atttgagtgc 420
caattcgatg atgagtcagc aactgtggtg gactttctga ggagatggat agccttctgt 480
caaagcatca tctcaacaag ccctcaaggt ggaggaggtt ctggaggcgg tggaagtggt 540
ggcggaggta gcagggtcat tccagtctct ggacctgcca ggtgtcttag ccagtcccga 600
aacctgctga agaccacaga tgacatggtg aagacggcca gagaaaaact gaaacattat 660
tcctgcactg ctgaagacat cgatcatgaa gacatcacac gggaccaaac cagcacattg 720
aagacctgtt taccactgga actacacaag aacgagagtt gcctggctac tagagagact 780
tcttccacaa caagagggag ctgcctgccc ccacagaaga cgtctttgat gatgaccctg 840
tgccttggta gcatctatga ggacttgaag atgtaccaga cagagttcca ggccatcaac 900
gcagcacttc agaatcacaa ccatcagcag atcattctag acaagggcat gctggtggcc 960
atcgatgagc tgatgcagtc tctgaatcat aatggcgaga ctctgcgcca gaaacctcct 1020
gtgggagaag cagaccctta cagagtgaaa atgaagctct gcatcctgct tcacgccttc 1080
agcacccgcg tcgtgaccat caacagggtg atgggctatc tgagctccgc cgagggcaga 1140
ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctatgtg gctgcagaat 1200
ttacttttcc tgggcattgt ggtctacagc ctctcagcac ccacccgctc acccatcact 1260
gtcacccggc cttggaagca tgtagaggcc atcaaagaag ccctgaacct cctggatgac 1320
atgcctgtca cgttgaatga agaggtagaa gtcgtctcta acgagttctc cttcaagaag 1380
ctaacatgtg tgcagacccg cctgaagata ttcgagcagg gtctacgggg caatttcacc 1440
aaactcaagg gcgccttgaa catgacagcc agctactacc agacatactg ccccccaact 1500
ccggaaacgg actgtgaaac acaagttacc acctatgcgg atttcataga cagccttaaa 1560
acctttctga ctgatatccc ctttgaatgc aaaaaaccag gccaaaaagg aggcggtgga 1620
agtggcggtg gaggctctgg aggtggcgga agcatgtggg agctggagaa agacgtttat 1680
gttgtagagg tggactggac tcccgatgcc cctggagaaa cagtgaacct cacctgtgac 1740
acgcctgaag aagatgacat cacctggacc tcagaccaga gacatggagt cataggctct 1800
ggaaagaccc tgaccatcac tgtcaaagag tttctagatg ctggccagta cacctgccac 1860
aaaggaggcg agactctgag ccactcacat ctgctgctcc acaagaagga aaatggaatt 1920
tggtccactg aaattttaaa aaatttcaaa aacaagactt tcctgaagtg tgaagcacca 1980
aattactccg gacggttcac gtgctcatgg ctggtgcaaa gaaacatgga cttgaagttc 2040
aacatcaaga gcagtagcag ttcccctgac tctcgggcag tgacatgtgg aatggcgtct 2100
ctgtctgcag agaaggtcac actggaccaa agggactatg agaagtattc agtgtcctgc 2160
caggaggatg tcacctgccc aactgccgag gagaccctgc ccattgaact ggcgttggaa 2220
gcacggcagc agaataaata tgagaactac agcaccagct tcttcatcag ggacatcatc 2280
aaaccagacc cgcccaagaa cttgcagatg aagcctttga agaactcaca ggtggaggtc 2340
agctgggagt accctgactc ctggagcact ccccattcct acttctccct caagttcttt 2400
gttcgaatcc agcgcaagaa agaaaagatg aaggagacag aggaggggtg taaccagaaa 2460
ggtgcgttcc tcgtagagaa gacatctacc gaagtccaat gcaaaggcgg gaatgtctgc 2520
gtgcaagctc aggatcgcta ttacaattcc tcgtgcagca agtgggcatg tgttccctgc 2580
agggtccgat cctag 2595
<210> 42
<211> 2616
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aGMCSFIL12bIL2蛋白质异二聚体的核苷酸序列
<400> 42
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc gcacccaccc gctcacccat cactgtcacc 720
cggccttgga agcatgtaga ggccatcaaa gaagccctga acctcctgga tgacatgcct 780
gtcacgttga atgaagaggt agaagtcgtc tctaacgagt tctccttcaa gaagctaaca 840
tgtgtgcaga cccgcctgaa gatattcgag cagggtctac ggggcaattt caccaaactc 900
aagggcgcct tgaacatgac agccagctac taccagacat actgcccccc aactccggaa 960
acggactgtg aaacacaagt taccacctat gcggatttca tagacagcct taaaaccttt 1020
ctgactgata tcccctttga atgcaaaaaa ccaggccaaa aagagggcag aggaagtctt 1080
ctaacatgcg gtgacgtgga ggagaatccc ggccctatgt gtcctcagaa gctaaccatc 1140
tcctggtttg ccatcgtttt gctggtgtct ccactcatgg ccatgtggga gctggagaaa 1200
gacgtttatg ttgtagaggt ggactggact cccgatgccc ctggagaaac agtgaacctc 1260
acctgtgaca cgcctgaaga agatgacatc acctggacct cagaccagag acatggagtc 1320
ataggctctg gaaagaccct gaccatcact gtcaaagagt ttctagatgc tggccagtac 1380
acctgccaca aaggaggcga gactctgagc cactcacatc tgctgctcca caagaaggaa 1440
aatggaattt ggtccactga aattttaaaa aatttcaaaa acaagacttt cctgaagtgt 1500
gaagcaccaa attactccgg acggttcacg tgctcatggc tggtgcaaag aaacatggac 1560
ttgaagttca acatcaagag cagtagcagt tcccctgact ctcgggcagt gacatgtgga 1620
atggcgtctc tgtctgcaga gaaggtcaca ctggaccaaa gggactatga gaagtattca 1680
gtgtcctgcc aggaggatgt cacctgccca actgccgagg agaccctgcc cattgaactg 1740
gcgttggaag cacggcagca gaataaatat gagaactaca gcaccagctt cttcatcagg 1800
gacatcatca aaccagaccc gcccaagaac ttgcagatga agcctttgaa gaactcacag 1860
gtggaggtca gctgggagta ccctgactcc tggagcactc cccattccta cttctccctc 1920
aagttctttg ttcgaatcca gcgcaagaaa gaaaagatga aggagacaga ggaggggtgt 1980
aaccagaaag gtgcgttcct cgtagagaag acatctaccg aagtccaatg caaaggcggg 2040
aatgtctgcg tgcaagctca ggatcgctat tacaattcct cgtgcagcaa gtgggcatgt 2100
gttccctgca gggtccgatc cggaggcggt ggaagtggcg gtggaggctc tggaggtggc 2160
ggaagcgcac ccacttcaag ctccacttca agctctacag cggaagcaca gcagcagcag 2220
cagcagcagc agcagcagca gcagcacctg gagcagctgt tgatggacct acaggagctc 2280
ctgagcagga tggagaatta caggaacctg aaactcccca ggatgctcac cttcaaattt 2340
tacttgccca agcaggccac agaattgaaa gatcttcagt gcctagaaga tgaacttgga 2400
cctctgcggc atgttctgga tttgactcaa agcaaaagct ttcaattgga agatgctgag 2460
aatttcatca gcaatatcag agtaactgtt gtaaaactaa agggctctga caacacattt 2520
gagtgccaat tcgatgatga gtcagcaact gtggtggact ttctgaggag atggatagcc 2580
ttctgtcaaa gcatcatctc aacaagccct caataa 2616
<210> 43
<211> 2556
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL7IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 43
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc gagtgccaca ttaaagacaa agaaggtaaa 720
gcatatgaga gtgtactgat gatcagcatc gatgaattgg acaaaatgac aggaactgat 780
agtaattgcc cgaataatga accaaacttt tttagaaaac atgtatgtga tgatacaaag 840
gaagctgctt ttctaaatcg tgctgctcgc aagttgaagc aatttcttaa aatgaatatc 900
agtgaagaat tcaatgtcca cttactaaca gtatcacaag gcacacaaac actggtgaac 960
tgcacaagta aggaagaaaa aaacgtaaag gaacagaaaa agaatgatgc atgtttccta 1020
aagagactac tgagagaaat aaaaacttgt tggaataaaa ttttgaaggg cagtatagag 1080
ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtgtcct 1140
cagaagctaa ccatctcctg gtttgccatc gttttgctgg tgtctccact catggccatg 1200
tgggagctgg agaaagacgt ttatgttgta gaggtggact ggactcccga tgcccctgga 1260
gaaacagtga acctcacctg tgacacgcct gaagaagatg acatcacctg gacctcagac 1320
cagagacatg gagtcatagg ctctggaaag accctgacca tcactgtcaa agagtttcta 1380
gatgctggcc agtacacctg ccacaaagga ggcgagactc tgagccactc acatctgctg 1440
ctccacaaga aggaaaatgg aatttggtcc actgaaattt taaaaaattt caaaaacaag 1500
actttcctga agtgtgaagc accaaattac tccggacggt tcacgtgctc atggctggtg 1560
caaagaaaca tggacttgaa gttcaacatc aagagcagta gcagttcccc tgactctcgg 1620
gcagtgacat gtggaatggc gtctctgtct gcagagaagg tcacactgga ccaaagggac 1680
tatgagaagt attcagtgtc ctgccaggag gatgtcacct gcccaactgc cgaggagacc 1740
ctgcccattg aactggcgtt ggaagcacgg cagcagaata aatatgagaa ctacagcacc 1800
agcttcttca tcagggacat catcaaacca gacccgccca agaacttgca gatgaagcct 1860
ttgaagaact cacaggtgga ggtcagctgg gagtaccctg actcctggag cactccccat 1920
tcctacttct ccctcaagtt ctttgttcga atccagcgca agaaagaaaa gatgaaggag 1980
acagaggagg ggtgtaacca gaaaggtgcg ttcctcgtag agaagacatc taccgaagtc 2040
caatgcaaag gcgggaatgt ctgcgtgcaa gctcaggatc gctattacaa ttcctcgtgc 2100
agcaagtggg catgtgttcc ctgcagggtc cgatccggag gcggtggaag tggcggtgga 2160
ggctctggag gtggcggaag cgcacccacc cgctcaccca tcactgtcac ccggccttgg 2220
aagcatgtag aggccatcaa agaagccctg aacctcctgg atgacatgcc tgtcacgttg 2280
aatgaagagg tagaagtcgt ctctaacgag ttctccttca agaagctaac atgtgtgcag 2340
acccgcctga agatattcga gcagggtcta cggggcaatt tcaccaaact caagggcgcc 2400
ttgaacatga cagccagcta ctaccagaca tactgccccc caactccgga aacggactgt 2460
gaaacacaag ttaccaccta tgcggatttc atagacagcc ttaaaacctt tctgactgat 2520
atcccctttg aatgcaaaaa accaggccaa aaatga 2556
<210> 44
<211> 2568
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL15IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 44
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc ggcattcatg tcttcatttt gggctgtgtc 720
agtgtaggtc tccctaaaac agaggccaac tggatagatg taagatatga cctggagaaa 780
attgaaagcc ttattcaatc tattcatatt gacaccactt tatacactga cagtgacttt 840
catcccagtt gcaaagttac tgcaatgaac tgctttctcc tggaattgca ggttatttta 900
catgagtaca gtaacatgac tcttaatgaa acagtaagaa acgtgctcta ccttgcaaac 960
agcactctgt cttctaacaa gaatgtagca gaatctggct gcaaggaatg tgaggagctg 1020
gaggagaaaa ccttcacaga gtttttgcaa agctttatac gcattgtcca aatgttcatc 1080
aacacgtccg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 1140
cctatgtgtc ctcagaagct aaccatctcc tggtttgcca tcgttttgct ggtgtctcca 1200
ctcatggcca tgtgggagct ggagaaagac gtttatgttg tagaggtgga ctggactccc 1260
gatgcccctg gagaaacagt gaacctcacc tgtgacacgc ctgaagaaga tgacatcacc 1320
tggacctcag accagagaca tggagtcata ggctctggaa agaccctgac catcactgtc 1380
aaagagtttc tagatgctgg ccagtacacc tgccacaaag gaggcgagac tctgagccac 1440
tcacatctgc tgctccacaa gaaggaaaat ggaatttggt ccactgaaat tttaaaaaat 1500
ttcaaaaaca agactttcct gaagtgtgaa gcaccaaatt actccggacg gttcacgtgc 1560
tcatggctgg tgcaaagaaa catggacttg aagttcaaca tcaagagcag tagcagttcc 1620
cctgactctc gggcagtgac atgtggaatg gcgtctctgt ctgcagagaa ggtcacactg 1680
gaccaaaggg actatgagaa gtattcagtg tcctgccagg aggatgtcac ctgcccaact 1740
gccgaggaga ccctgcccat tgaactggcg ttggaagcac ggcagcagaa taaatatgag 1800
aactacagca ccagcttctt catcagggac atcatcaaac cagacccgcc caagaacttg 1860
cagatgaagc ctttgaagaa ctcacaggtg gaggtcagct gggagtaccc tgactcctgg 1920
agcactcccc attcctactt ctccctcaag ttctttgttc gaatccagcg caagaaagaa 1980
aagatgaagg agacagagga ggggtgtaac cagaaaggtg cgttcctcgt agagaagaca 2040
tctaccgaag tccaatgcaa aggcgggaat gtctgcgtgc aagctcagga tcgctattac 2100
aattcctcgt gcagcaagtg ggcatgtgtt ccctgcaggg tccgatccgg aggcggtgga 2160
agtggcggtg gaggctctgg aggtggcgga agcgcaccca cccgctcacc catcactgtc 2220
acccggcctt ggaagcatgt agaggccatc aaagaagccc tgaacctcct ggatgacatg 2280
cctgtcacgt tgaatgaaga ggtagaagtc gtctctaacg agttctcctt caagaagcta 2340
acatgtgtgc agacccgcct gaagatattc gagcagggtc tacggggcaa tttcaccaaa 2400
ctcaagggcg ccttgaacat gacagccagc tactaccaga catactgccc cccaactccg 2460
gaaacggact gtgaaacaca agttaccacc tatgcggatt tcatagacag ccttaaaacc 2520
tttctgactg atatcccctt tgaatgcaaa aaaccaggcc aaaaatga 2568
<210> 45
<211> 2556
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL21IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 45
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc cataaatcaa gcccccaagg gccagatcgc 720
ctcctgatta gacttcgtca ccttattgac attgttgaac agctgaaaat ctatgaaaat 780
gacttggatc ctgaacttct atcagctcca caagatgtaa aggggcactg tgagcatgca 840
gcttttgcct gttttcagaa ggccaaactc aagccatcaa accctggaaa caataagaca 900
ttcatcattg acctcgtggc ccagctcagg aggaggctgc ctgccaggag gggaggaaag 960
aaacagaagc acatagctaa atgcccttcc tgtgattcgt atgagaaaag gacacccaaa 1020
gaattcctag aaagactaaa atggctcctt caaaagatga ttcatcagca tctctccgag 1080
ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtgtcct 1140
cagaagctaa ccatctcctg gtttgccatc gttttgctgg tgtctccact catggccatg 1200
tgggagctgg agaaagacgt ttatgttgta gaggtggact ggactcccga tgcccctgga 1260
gaaacagtga acctcacctg tgacacgcct gaagaagatg acatcacctg gacctcagac 1320
cagagacatg gagtcatagg ctctggaaag accctgacca tcactgtcaa agagtttcta 1380
gatgctggcc agtacacctg ccacaaagga ggcgagactc tgagccactc acatctgctg 1440
ctccacaaga aggaaaatgg aatttggtcc actgaaattt taaaaaattt caaaaacaag 1500
actttcctga agtgtgaagc accaaattac tccggacggt tcacgtgctc atggctggtg 1560
caaagaaaca tggacttgaa gttcaacatc aagagcagta gcagttcccc tgactctcgg 1620
gcagtgacat gtggaatggc gtctctgtct gcagagaagg tcacactgga ccaaagggac 1680
tatgagaagt attcagtgtc ctgccaggag gatgtcacct gcccaactgc cgaggagacc 1740
ctgcccattg aactggcgtt ggaagcacgg cagcagaata aatatgagaa ctacagcacc 1800
agcttcttca tcagggacat catcaaacca gacccgccca agaacttgca gatgaagcct 1860
ttgaagaact cacaggtgga ggtcagctgg gagtaccctg actcctggag cactccccat 1920
tcctacttct ccctcaagtt ctttgttcga atccagcgca agaaagaaaa gatgaaggag 1980
acagaggagg ggtgtaacca gaaaggtgcg ttcctcgtag agaagacatc taccgaagtc 2040
caatgcaaag gcgggaatgt ctgcgtgcaa gctcaggatc gctattacaa ttcctcgtgc 2100
agcaagtggg catgtgttcc ctgcagggtc cgatccggag gcggtggaag tggcggtgga 2160
ggctctggag gtggcggaag cgcacccacc cgctcaccca tcactgtcac ccggccttgg 2220
aagcatgtag aggccatcaa agaagccctg aacctcctgg atgacatgcc tgtcacgttg 2280
aatgaagagg tagaagtcgt ctctaacgag ttctccttca agaagctaac atgtgtgcag 2340
acccgcctga agatattcga gcagggtcta cggggcaatt tcaccaaact caagggcgcc 2400
ttgaacatga cagccagcta ctaccagaca tactgccccc caactccgga aacggactgt 2460
gaaacacaag ttaccaccta tgcggatttc atagacagcc ttaaaacctt tctgactgat 2520
atcccctttg aatgcaaaaa accaggccaa aaatga 2556
<210> 46
<211> 2862
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL2IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 46
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc gcacccactt caagctccac ttcaagctct 720
acagcggaag cacagcagca gcagcagcag cagcagcagc agcagcagca cctggagcag 780
ctgttgatgg acctacagga gctcctgagc aggatggaga attacaggaa cctgaaactc 840
cccaggatgc tcaccttcaa attttacttg cccaagcagg ccacagaatt gaaagatctt 900
cagtgcctag aagatgaact tggacctctg cggcatgttc tggatttgac tcaaagcaaa 960
agctttcaat tggaagatgc tgagaatttc atcagcaata tcagagtaac tgttgtaaaa 1020
ctaaagggct ctgacaacac atttgagtgc caattcgatg atgagtcagc aactgtggtg 1080
gactttctga ggagatggat agccttctgt caaagcatca tctcaacaag ccctcaagag 1140
ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtgtcct 1200
cagaagctaa ccatctcctg gtttgccatc gttttgctgg tgtctccact catggccatg 1260
tgggagctgg agaaagacgt ttatgttgta gaggtggact ggactcccga tgcccctgga 1320
gaaacagtga acctcacctg tgacacgcct gaagaagatg acatcacctg gacctcagac 1380
cagagacatg gagtcatagg ctctggaaag accctgacca tcactgtcaa agagtttcta 1440
gatgctggcc agtacacctg ccacaaagga ggcgagactc tgagccactc acatctgctg 1500
ctccacaaga aggaaaatgg aatttggtcc actgaaattt taaaaaattt caaaaacaag 1560
actttcctga agtgtgaagc accaaattac tccggacggt tcacgtgctc atggctggtg 1620
caaagaaaca tggacttgaa gttcaacatc aagagcagta gcagttcccc tgactctcgg 1680
gcagtgacat gtggaatggc gtctctgtct gcagagaagg tcacactgga ccaaagggac 1740
tatgagaagt attcagtgtc ctgccaggag gatgtcacct gcccaactgc cgaggagacc 1800
ctgcccattg aactggcgtt ggaagcacgg cagcagaata aatatgagaa ctacagcacc 1860
agcttcttca tcagggacat catcaaacca gacccgccca agaacttgca gatgaagcct 1920
ttgaagaact cacaggtgga ggtcagctgg gagtaccctg actcctggag cactccccat 1980
tcctacttct ccctcaagtt ctttgttcga atccagcgca agaaagaaaa gatgaaggag 2040
acagaggagg ggtgtaacca gaaaggtgcg ttcctcgtag agaagacatc taccgaagtc 2100
caatgcaaag gcgggaatgt ctgcgtgcaa gctcaggatc gctattacaa ttcctcgtgc 2160
agcaagtggg catgtgttcc ctgcagggtc cgatccggag gcggtggaag tggcggtgga 2220
ggctctggag gtggcggaag cgggacacct gactgttact tcagccacag tcccatctcc 2280
tccaacttca aagtgaagtt tagagagttg actgaccacc tgcttaaaga ttacccagtc 2340
actgtggccg tcaatcttca ggacgagaag cactgcaagg ccttgtggag cctcttccta 2400
gcccagcgct ggatagagca actgaagact gtggcagggt ctaagatgca aacgcttctg 2460
gaggacgtca acaccgagat acattttgtc acctcatgta ccttccagcc cctaccagaa 2520
tgtctgcgat tcgtccagac caacatctcc cacctcctga aggacacctg cacacagctg 2580
cttgctctga agccctgtat cgggaaggcc tgccagaatt tctctcggtg cctggaggtg 2640
cagtgccagc cggactcctc caccctgctg cccccaagga gtcccatagc cctagaagcc 2700
acggagctcc cagagcctcg gcccaggcag ctgttgctcc tgctgctgct gctgctgcct 2760
ctcacactgg tgctgctggc agccgcctgg ggccttcgct ggcaaagggc aagaaggagg 2820
ggggagctcc accctggggt gcccctcccc tcccatccct ag 2862
<210> 47
<211> 2802
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL7IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 47
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc gagtgccaca ttaaagacaa agaaggtaaa 720
gcatatgaga gtgtactgat gatcagcatc gatgaattgg acaaaatgac aggaactgat 780
agtaattgcc cgaataatga accaaacttt tttagaaaac atgtatgtga tgatacaaag 840
gaagctgctt ttctaaatcg tgctgctcgc aagttgaagc aatttcttaa aatgaatatc 900
agtgaagaat tcaatgtcca cttactaaca gtatcacaag gcacacaaac actggtgaac 960
tgcacaagta aggaagaaaa aaacgtaaag gaacagaaaa agaatgatgc atgtttccta 1020
aagagactac tgagagaaat aaaaacttgt tggaataaaa ttttgaaggg cagtatagag 1080
ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtgtcct 1140
cagaagctaa ccatctcctg gtttgccatc gttttgctgg tgtctccact catggccatg 1200
tgggagctgg agaaagacgt ttatgttgta gaggtggact ggactcccga tgcccctgga 1260
gaaacagtga acctcacctg tgacacgcct gaagaagatg acatcacctg gacctcagac 1320
cagagacatg gagtcatagg ctctggaaag accctgacca tcactgtcaa agagtttcta 1380
gatgctggcc agtacacctg ccacaaagga ggcgagactc tgagccactc acatctgctg 1440
ctccacaaga aggaaaatgg aatttggtcc actgaaattt taaaaaattt caaaaacaag 1500
actttcctga agtgtgaagc accaaattac tccggacggt tcacgtgctc atggctggtg 1560
caaagaaaca tggacttgaa gttcaacatc aagagcagta gcagttcccc tgactctcgg 1620
gcagtgacat gtggaatggc gtctctgtct gcagagaagg tcacactgga ccaaagggac 1680
tatgagaagt attcagtgtc ctgccaggag gatgtcacct gcccaactgc cgaggagacc 1740
ctgcccattg aactggcgtt ggaagcacgg cagcagaata aatatgagaa ctacagcacc 1800
agcttcttca tcagggacat catcaaacca gacccgccca agaacttgca gatgaagcct 1860
ttgaagaact cacaggtgga ggtcagctgg gagtaccctg actcctggag cactccccat 1920
tcctacttct ccctcaagtt ctttgttcga atccagcgca agaaagaaaa gatgaaggag 1980
acagaggagg ggtgtaacca gaaaggtgcg ttcctcgtag agaagacatc taccgaagtc 2040
caatgcaaag gcgggaatgt ctgcgtgcaa gctcaggatc gctattacaa ttcctcgtgc 2100
agcaagtggg catgtgttcc ctgcagggtc cgatccggag gcggtggaag tggcggtgga 2160
ggctctggag gtggcggaag cgggacacct gactgttact tcagccacag tcccatctcc 2220
tccaacttca aagtgaagtt tagagagttg actgaccacc tgcttaaaga ttacccagtc 2280
actgtggccg tcaatcttca ggacgagaag cactgcaagg ccttgtggag cctcttccta 2340
gcccagcgct ggatagagca actgaagact gtggcagggt ctaagatgca aacgcttctg 2400
gaggacgtca acaccgagat acattttgtc acctcatgta ccttccagcc cctaccagaa 2460
tgtctgcgat tcgtccagac caacatctcc cacctcctga aggacacctg cacacagctg 2520
cttgctctga agccctgtat cgggaaggcc tgccagaatt tctctcggtg cctggaggtg 2580
cagtgccagc cggactcctc caccctgctg cccccaagga gtcccatagc cctagaagcc 2640
acggagctcc cagagcctcg gcccaggcag ctgttgctcc tgctgctgct gctgctgcct 2700
ctcacactgg tgctgctggc agccgcctgg ggccttcgct ggcaaagggc aagaaggagg 2760
ggggagctcc accctggggt gcccctcccc tcccatccct ag 2802
<210> 48
<211> 2814
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL15IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 48
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc ggcattcatg tcttcatttt gggctgtgtc 720
agtgtaggtc tccctaaaac agaggccaac tggatagatg taagatatga cctggagaaa 780
attgaaagcc ttattcaatc tattcatatt gacaccactt tatacactga cagtgacttt 840
catcccagtt gcaaagttac tgcaatgaac tgctttctcc tggaattgca ggttatttta 900
catgagtaca gtaacatgac tcttaatgaa acagtaagaa acgtgctcta ccttgcaaac 960
agcactctgt cttctaacaa gaatgtagca gaatctggct gcaaggaatg tgaggagctg 1020
gaggagaaaa ccttcacaga gtttttgcaa agctttatac gcattgtcca aatgttcatc 1080
aacacgtccg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc 1140
cctatgtgtc ctcagaagct aaccatctcc tggtttgcca tcgttttgct ggtgtctcca 1200
ctcatggcca tgtgggagct ggagaaagac gtttatgttg tagaggtgga ctggactccc 1260
gatgcccctg gagaaacagt gaacctcacc tgtgacacgc ctgaagaaga tgacatcacc 1320
tggacctcag accagagaca tggagtcata ggctctggaa agaccctgac catcactgtc 1380
aaagagtttc tagatgctgg ccagtacacc tgccacaaag gaggcgagac tctgagccac 1440
tcacatctgc tgctccacaa gaaggaaaat ggaatttggt ccactgaaat tttaaaaaat 1500
ttcaaaaaca agactttcct gaagtgtgaa gcaccaaatt actccggacg gttcacgtgc 1560
tcatggctgg tgcaaagaaa catggacttg aagttcaaca tcaagagcag tagcagttcc 1620
cctgactctc gggcagtgac atgtggaatg gcgtctctgt ctgcagagaa ggtcacactg 1680
gaccaaaggg actatgagaa gtattcagtg tcctgccagg aggatgtcac ctgcccaact 1740
gccgaggaga ccctgcccat tgaactggcg ttggaagcac ggcagcagaa taaatatgag 1800
aactacagca ccagcttctt catcagggac atcatcaaac cagacccgcc caagaacttg 1860
cagatgaagc ctttgaagaa ctcacaggtg gaggtcagct gggagtaccc tgactcctgg 1920
agcactcccc attcctactt ctccctcaag ttctttgttc gaatccagcg caagaaagaa 1980
aagatgaagg agacagagga ggggtgtaac cagaaaggtg cgttcctcgt agagaagaca 2040
tctaccgaag tccaatgcaa aggcgggaat gtctgcgtgc aagctcagga tcgctattac 2100
aattcctcgt gcagcaagtg ggcatgtgtt ccctgcaggg tccgatccgg aggcggtgga 2160
agtggcggtg gaggctctgg aggtggcgga agcgggacac ctgactgtta cttcagccac 2220
agtcccatct cctccaactt caaagtgaag tttagagagt tgactgacca cctgcttaaa 2280
gattacccag tcactgtggc cgtcaatctt caggacgaga agcactgcaa ggccttgtgg 2340
agcctcttcc tagcccagcg ctggatagag caactgaaga ctgtggcagg gtctaagatg 2400
caaacgcttc tggaggacgt caacaccgag atacattttg tcacctcatg taccttccag 2460
cccctaccag aatgtctgcg attcgtccag accaacatct cccacctcct gaaggacacc 2520
tgcacacagc tgcttgctct gaagccctgt atcgggaagg cctgccagaa tttctctcgg 2580
tgcctggagg tgcagtgcca gccggactcc tccaccctgc tgcccccaag gagtcccata 2640
gccctagaag ccacggagct cccagagcct cggcccaggc agctgttgct cctgctgctg 2700
ctgctgctgc ctctcacact ggtgctgctg gcagccgcct ggggccttcg ctggcaaagg 2760
gcaagaagga ggggggagct ccaccctggg gtgcccctcc cctcccatcc ctag 2814
<210> 49
<211> 2802
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码mIL12aIL21IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 49
atgtgtcaat cacgctacct cctctttttg gccacccttg ccctcctaaa ccacctcagt 60
ttggccaggg tcattccagt ctctggacct gccaggtgtc ttagccagtc ccgaaacctg 120
ctgaagacca cagatgacat ggtgaagacg gccagagaaa aactgaaaca ttattcctgc 180
actgctgaag acatcgatca tgaagacatc acacgggacc aaaccagcac attgaagacc 240
tgtttaccac tggaactaca caagaacgag agttgcctgg ctactagaga gacttcttcc 300
acaacaagag ggagctgcct gcccccacag aagacgtctt tgatgatgac cctgtgcctt 360
ggtagcatct atgaggactt gaagatgtac cagacagagt tccaggccat caacgcagca 420
cttcagaatc acaaccatca gcagatcatt ctagacaagg gcatgctggt ggccatcgat 480
gagctgatgc agtctctgaa tcataatggc gagactctgc gccagaaacc tcctgtggga 540
gaagcagacc cttacagagt gaaaatgaag ctctgcatcc tgcttcacgc cttcagcacc 600
cgcgtcgtga ccatcaacag ggtgatgggc tatctgagct ccgccggtgg aggaggttct 660
ggaggcggtg gaagtggtgg cggaggtagc cataaatcaa gcccccaagg gccagatcgc 720
ctcctgatta gacttcgtca ccttattgac attgttgaac agctgaaaat ctatgaaaat 780
gacttggatc ctgaacttct atcagctcca caagatgtaa aggggcactg tgagcatgca 840
gcttttgcct gttttcagaa ggccaaactc aagccatcaa accctggaaa caataagaca 900
ttcatcattg acctcgtggc ccagctcagg aggaggctgc ctgccaggag gggaggaaag 960
aaacagaagc acatagctaa atgcccttcc tgtgattcgt atgagaaaag gacacccaaa 1020
gaattcctag aaagactaaa atggctcctt caaaagatga ttcatcagca tctctccgag 1080
ggcagaggaa gtcttctaac atgcggtgac gtggaggaga atcccggccc tatgtgtcct 1140
cagaagctaa ccatctcctg gtttgccatc gttttgctgg tgtctccact catggccatg 1200
tgggagctgg agaaagacgt ttatgttgta gaggtggact ggactcccga tgcccctgga 1260
gaaacagtga acctcacctg tgacacgcct gaagaagatg acatcacctg gacctcagac 1320
cagagacatg gagtcatagg ctctggaaag accctgacca tcactgtcaa agagtttcta 1380
gatgctggcc agtacacctg ccacaaagga ggcgagactc tgagccactc acatctgctg 1440
ctccacaaga aggaaaatgg aatttggtcc actgaaattt taaaaaattt caaaaacaag 1500
actttcctga agtgtgaagc accaaattac tccggacggt tcacgtgctc atggctggtg 1560
caaagaaaca tggacttgaa gttcaacatc aagagcagta gcagttcccc tgactctcgg 1620
gcagtgacat gtggaatggc gtctctgtct gcagagaagg tcacactgga ccaaagggac 1680
tatgagaagt attcagtgtc ctgccaggag gatgtcacct gcccaactgc cgaggagacc 1740
ctgcccattg aactggcgtt ggaagcacgg cagcagaata aatatgagaa ctacagcacc 1800
agcttcttca tcagggacat catcaaacca gacccgccca agaacttgca gatgaagcct 1860
ttgaagaact cacaggtgga ggtcagctgg gagtaccctg actcctggag cactccccat 1920
tcctacttct ccctcaagtt ctttgttcga atccagcgca agaaagaaaa gatgaaggag 1980
acagaggagg ggtgtaacca gaaaggtgcg ttcctcgtag agaagacatc taccgaagtc 2040
caatgcaaag gcgggaatgt ctgcgtgcaa gctcaggatc gctattacaa ttcctcgtgc 2100
agcaagtggg catgtgttcc ctgcagggtc cgatccggag gcggtggaag tggcggtgga 2160
ggctctggag gtggcggaag cgggacacct gactgttact tcagccacag tcccatctcc 2220
tccaacttca aagtgaagtt tagagagttg actgaccacc tgcttaaaga ttacccagtc 2280
actgtggccg tcaatcttca ggacgagaag cactgcaagg ccttgtggag cctcttccta 2340
gcccagcgct ggatagagca actgaagact gtggcagggt ctaagatgca aacgcttctg 2400
gaggacgtca acaccgagat acattttgtc acctcatgta ccttccagcc cctaccagaa 2460
tgtctgcgat tcgtccagac caacatctcc cacctcctga aggacacctg cacacagctg 2520
cttgctctga agccctgtat cgggaaggcc tgccagaatt tctctcggtg cctggaggtg 2580
cagtgccagc cggactcctc caccctgctg cccccaagga gtcccatagc cctagaagcc 2640
acggagctcc cagagcctcg gcccaggcag ctgttgctcc tgctgctgct gctgctgcct 2700
ctcacactgg tgctgctggc agccgcctgg ggccttcgct ggcaaagggc aagaaggagg 2760
ggggagctcc accctggggt gcccctcccc tcccatccct ag 2802
<210> 50
<211> 2670
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL2IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 50
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagcgcacct acttcaagtt ctacaaagaa aacacagcta 840
caactggagc atttactgct ggatttacag atgattttga atggaattaa taattacaag 900
aatcccaaac tcaccaggat gctcacattt aagttttaca tgcccaagaa ggccacagaa 960
ctgaaacatc ttcagtgtct agaagaagaa ctcaaacctc tggaggaagt gctaaattta 1020
gctcaaagca aaaactttca cttaagaccc agggacttaa tcagcaatat caacgtaata 1080
gttctggaac taaagggatc tgaaacaaca ttcatgtgtg aatatgctga tgagacagca 1140
accattgtag aatttctgaa cagatggatt accttttgtc aaagcatcat ctcaacactg 1200
actgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 1260
tgtcaccagc agttggtcat ctcttggttt tccctggttt ttctggcatc tcccctcgtg 1320
gccatatggg aactgaagaa agatgtttat gtcgtagaat tggattggta tccggatgcc 1380
cctggagaaa tggtggtcct cacctgtgac acccctgaag aagatggtat cacctggacc 1440
ttggaccaga gcagtgaggt cttaggctct ggcaaaaccc tgaccatcca agtcaaagag 1500
tttggagatg ctggccagta cacctgtcac aaaggaggcg aggttctaag ccattcgctc 1560
ctgctgcttc acaaaaagga agatggaatt tggtccactg atattttaaa ggaccagaaa 1620
gaacccaaaa ataagacctt tctaagatgc gaggccaaga attattctgg acgtttcacc 1680
tgctggtggc tgacgacaat cagtactgat ttgacattca gtgtcaaaag cagcagaggc 1740
tcttctgacc cccaaggggt gacgtgcgga gctgctacac tctctgcaga gagagtcaga 1800
ggggacaaca aggagtatga gtactcagtg gagtgccagg aggacagtgc ctgcccagct 1860
gctgaggaga gtctgcccat tgaggtcatg gtggatgccg ttcacaagct caagtatgaa 1920
aactacacca gcagcttctt catcagggac atcatcaaac ctgacccacc caagaacttg 1980
cagctgaagc cattaaagaa ttctcggcag gtggaggtca gctgggagta ccctgacacc 2040
tggagtactc cacattccta cttctccctg acattctgcg ttcaggtcca gggcaagagc 2100
aagagagaaa agaaagatag agtcttcacg gacaagacct cagccacggt catctgccgc 2160
aaaaatgcca gcattagcgt gcgggcccag gaccgctact atagctcatc ttggagcgaa 2220
tgggcatctg tgccctgcag tggaggcggt ggaagtggcg gtggaggctc tggaggtggc 2280
ggaagcgcac ccgcccgctc gcccagcccc agcacgcagc cctgggagca tgtgaatgcc 2340
atccaggagg cccggcgtct cctgaacctg agtagagaca ctgctgctga gatgaatgaa 2400
acagtagaag tcatctcaga aatgtttgac ctccaggagc cgacctgcct acagacccgc 2460
ctggagctgt acaagcaggg cctgcggggc agcctcacca agctcaaggg ccccttgacc 2520
atgatggcca gccactacaa gcagcactgc cctccaaccc cggaaacttc ctgtgcaacc 2580
cagattatca cctttgaaag tttcaaagag aacctgaagg actttctgct tgtcatcccc 2640
tttgactgct gggagccagt ccaggagtga 2670
<210> 51
<211> 2727
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL7IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 51
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagcgattgt gatattgaag gtaaagatgg caaacaatat 840
gagagtgttc taatggtcag catcgatcaa ttattggaca gcatgaaaga aattggtagc 900
aattgcctga ataatgaatt taactttttt aaaagacata tctgtgatgc taataaggaa 960
ggtatgtttt tattccgtgc tgctcgcaag ttgaggcaat ttcttaaaat gaatagcact 1020
ggtgattttg atctccactt attaaaagtt tcagaaggca caacaatact gttgaactgc 1080
actggccagg ttaaaggaag aaaaccagct gccctgggtg aagcccaacc aacaaagagt 1140
ttggaagaaa ataaatcttt aaaggaacag aaaaaactga atgacttgtg tttcctaaag 1200
agactattac aagagataaa aacttgttgg aataaaattt tgatgggcac taaagaacac 1260
gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgtgt 1320
caccagcagt tggtcatctc ttggttttcc ctggtttttc tggcatctcc cctcgtggcc 1380
atatgggaac tgaagaaaga tgtttatgtc gtagaattgg attggtatcc ggatgcccct 1440
ggagaaatgg tggtcctcac ctgtgacacc cctgaagaag atggtatcac ctggaccttg 1500
gaccagagca gtgaggtctt aggctctggc aaaaccctga ccatccaagt caaagagttt 1560
ggagatgctg gccagtacac ctgtcacaaa ggaggcgagg ttctaagcca ttcgctcctg 1620
ctgcttcaca aaaaggaaga tggaatttgg tccactgata ttttaaagga ccagaaagaa 1680
cccaaaaata agacctttct aagatgcgag gccaagaatt attctggacg tttcacctgc 1740
tggtggctga cgacaatcag tactgatttg acattcagtg tcaaaagcag cagaggctct 1800
tctgaccccc aaggggtgac gtgcggagct gctacactct ctgcagagag agtcagaggg 1860
gacaacaagg agtatgagta ctcagtggag tgccaggagg acagtgcctg cccagctgct 1920
gaggagagtc tgcccattga ggtcatggtg gatgccgttc acaagctcaa gtatgaaaac 1980
tacaccagca gcttcttcat cagggacatc atcaaacctg acccacccaa gaacttgcag 2040
ctgaagccat taaagaattc tcggcaggtg gaggtcagct gggagtaccc tgacacctgg 2100
agtactccac attcctactt ctccctgaca ttctgcgttc aggtccaggg caagagcaag 2160
agagaaaaga aagatagagt cttcacggac aagacctcag ccacggtcat ctgccgcaaa 2220
aatgccagca ttagcgtgcg ggcccaggac cgctactata gctcatcttg gagcgaatgg 2280
gcatctgtgc cctgcagtgg aggcggtgga agtggcggtg gaggctctgg aggtggcgga 2340
agcgcacccg cccgctcgcc cagccccagc acgcagccct gggagcatgt gaatgccatc 2400
caggaggccc ggcgtctcct gaacctgagt agagacactg ctgctgagat gaatgaaaca 2460
gtagaagtca tctcagaaat gtttgacctc caggagccga cctgcctaca gacccgcctg 2520
gagctgtaca agcagggcct gcggggcagc ctcaccaagc tcaagggccc cttgaccatg 2580
atggccagcc actacaagca gcactgccct ccaaccccgg aaacttcctg tgcaacccag 2640
attatcacct ttgaaagttt caaagagaac ctgaaggact ttctgcttgt catccccttt 2700
gactgctggg agccagtcca ggagtga 2727
<210> 52
<211> 2670
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL15IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 52
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagcggcatt catgtcttca ttttgggctg tttcagtgca 840
gggcttccta aaacagaagc caactgggtg aatgtaataa gtgatttgaa aaaaattgaa 900
gatcttattc aatctatgca tattgatgct actttatata cggaaagtga tgttcacccc 960
agttgcaaag taacagcaat gaagtgcttt ctcttggagt tacaagttat ttcacttgag 1020
tccggagatg caagtattca tgatacagta gaaaatctga tcatcctagc aaacaacagt 1080
ttgtcttcta atgggaatgt aacagaatct ggatgcaaag aatgtgagga actggaggaa 1140
aaaaatatta aagaattttt gcagagtttt gtacatattg tccaaatgtt catcaacact 1200
tctgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 1260
tgtcaccagc agttggtcat ctcttggttt tccctggttt ttctggcatc tcccctcgtg 1320
gccatatggg aactgaagaa agatgtttat gtcgtagaat tggattggta tccggatgcc 1380
cctggagaaa tggtggtcct cacctgtgac acccctgaag aagatggtat cacctggacc 1440
ttggaccaga gcagtgaggt cttaggctct ggcaaaaccc tgaccatcca agtcaaagag 1500
tttggagatg ctggccagta cacctgtcac aaaggaggcg aggttctaag ccattcgctc 1560
ctgctgcttc acaaaaagga agatggaatt tggtccactg atattttaaa ggaccagaaa 1620
gaacccaaaa ataagacctt tctaagatgc gaggccaaga attattctgg acgtttcacc 1680
tgctggtggc tgacgacaat cagtactgat ttgacattca gtgtcaaaag cagcagaggc 1740
tcttctgacc cccaaggggt gacgtgcgga gctgctacac tctctgcaga gagagtcaga 1800
ggggacaaca aggagtatga gtactcagtg gagtgccagg aggacagtgc ctgcccagct 1860
gctgaggaga gtctgcccat tgaggtcatg gtggatgccg ttcacaagct caagtatgaa 1920
aactacacca gcagcttctt catcagggac atcatcaaac ctgacccacc caagaacttg 1980
cagctgaagc cattaaagaa ttctcggcag gtggaggtca gctgggagta ccctgacacc 2040
tggagtactc cacattccta cttctccctg acattctgcg ttcaggtcca gggcaagagc 2100
aagagagaaa agaaagatag agtcttcacg gacaagacct cagccacggt catctgccgc 2160
aaaaatgcca gcattagcgt gcgggcccag gaccgctact atagctcatc ttggagcgaa 2220
tgggcatctg tgccctgcag tggaggcggt ggaagtggcg gtggaggctc tggaggtggc 2280
ggaagcgcac ccgcccgctc gcccagcccc agcacgcagc cctgggagca tgtgaatgcc 2340
atccaggagg cccggcgtct cctgaacctg agtagagaca ctgctgctga gatgaatgaa 2400
acagtagaag tcatctcaga aatgtttgac ctccaggagc cgacctgcct acagacccgc 2460
ctggagctgt acaagcaggg cctgcggggc agcctcacca agctcaaggg ccccttgacc 2520
atgatggcca gccactacaa gcagcactgc cctccaaccc cggaaacttc ctgtgcaacc 2580
cagattatca cctttgaaag tttcaaagag aacctgaagg actttctgct tgtcatcccc 2640
tttgactgct gggagccagt ccaggagtga 2670
<210> 53
<211> 2670
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL21IL12bGMCSF蛋白质异二聚体的核苷酸序列
<400> 53
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagccaaggt caagatcgcc acatgattag aatgcgtcaa 840
cttatagata ttgttgatca gctgaaaaat tatgtgaatg acttggtccc tgaatttctg 900
ccagctccag aagatgtaga gacaaactgt gagtggtcag ctttttcctg ctttcagaag 960
gcccaactaa agtcagcaaa tacaggaaac aatgaaagga taatcaatgt atcaattaaa 1020
aagctgaaga ggaaaccacc ttccacaaat gcagggagaa gacagaaaca cagactaaca 1080
tgcccttcat gtgattctta tgagaaaaaa ccacccaaag aattcctaga aagattcaaa 1140
tcacttctcc aaaagatgat tcatcagcat ctgtcctcta gaacacacgg aagtgaagat 1200
tccgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 1260
tgtcaccagc agttggtcat ctcttggttt tccctggttt ttctggcatc tcccctcgtg 1320
gccatatggg aactgaagaa agatgtttat gtcgtagaat tggattggta tccggatgcc 1380
cctggagaaa tggtggtcct cacctgtgac acccctgaag aagatggtat cacctggacc 1440
ttggaccaga gcagtgaggt cttaggctct ggcaaaaccc tgaccatcca agtcaaagag 1500
tttggagatg ctggccagta cacctgtcac aaaggaggcg aggttctaag ccattcgctc 1560
ctgctgcttc acaaaaagga agatggaatt tggtccactg atattttaaa ggaccagaaa 1620
gaacccaaaa ataagacctt tctaagatgc gaggccaaga attattctgg acgtttcacc 1680
tgctggtggc tgacgacaat cagtactgat ttgacattca gtgtcaaaag cagcagaggc 1740
tcttctgacc cccaaggggt gacgtgcgga gctgctacac tctctgcaga gagagtcaga 1800
ggggacaaca aggagtatga gtactcagtg gagtgccagg aggacagtgc ctgcccagct 1860
gctgaggaga gtctgcccat tgaggtcatg gtggatgccg ttcacaagct caagtatgaa 1920
aactacacca gcagcttctt catcagggac atcatcaaac ctgacccacc caagaacttg 1980
cagctgaagc cattaaagaa ttctcggcag gtggaggtca gctgggagta ccctgacacc 2040
tggagtactc cacattccta cttctccctg acattctgcg ttcaggtcca gggcaagagc 2100
aagagagaaa agaaagatag agtcttcacg gacaagacct cagccacggt catctgccgc 2160
aaaaatgcca gcattagcgt gcgggcccag gaccgctact atagctcatc ttggagcgaa 2220
tgggcatctg tgccctgcag tggaggcggt ggaagtggcg gtggaggctc tggaggtggc 2280
ggaagcgcac ccgcccgctc gcccagcccc agcacgcagc cctgggagca tgtgaatgcc 2340
atccaggagg cccggcgtct cctgaacctg agtagagaca ctgctgctga gatgaatgaa 2400
acagtagaag tcatctcaga aatgtttgac ctccaggagc cgacctgcct acagacccgc 2460
ctggagctgt acaagcaggg cctgcggggc agcctcacca agctcaaggg ccccttgacc 2520
atgatggcca gccactacaa gcagcactgc cctccaaccc cggaaacttc ctgtgcaacc 2580
cagattatca cctttgaaag tttcaaagag aacctgaagg actttctgct tgtcatcccc 2640
tttgactgct gggagccagt ccaggagtga 2670
<210> 54
<211> 2916
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL2IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 54
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagcgcacct acttcaagtt ctacaaagaa aacacagcta 840
caactggagc atttactgct ggatttacag atgattttga atggaattaa taattacaag 900
aatcccaaac tcaccaggat gctcacattt aagttttaca tgcccaagaa ggccacagaa 960
ctgaaacatc ttcagtgtct agaagaagaa ctcaaacctc tggaggaagt gctaaattta 1020
gctcaaagca aaaactttca cttaagaccc agggacttaa tcagcaatat caacgtaata 1080
gttctggaac taaagggatc tgaaacaaca ttcatgtgtg aatatgctga tgagacagca 1140
accattgtag aatttctgaa cagatggatt accttttgtc aaagcatcat ctcaacactg 1200
actgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 1260
tgtcaccagc agttggtcat ctcttggttt tccctggttt ttctggcatc tcccctcgtg 1320
gccatatggg aactgaagaa agatgtttat gtcgtagaat tggattggta tccggatgcc 1380
cctggagaaa tggtggtcct cacctgtgac acccctgaag aagatggtat cacctggacc 1440
ttggaccaga gcagtgaggt cttaggctct ggcaaaaccc tgaccatcca agtcaaagag 1500
tttggagatg ctggccagta cacctgtcac aaaggaggcg aggttctaag ccattcgctc 1560
ctgctgcttc acaaaaagga agatggaatt tggtccactg atattttaaa ggaccagaaa 1620
gaacccaaaa ataagacctt tctaagatgc gaggccaaga attattctgg acgtttcacc 1680
tgctggtggc tgacgacaat cagtactgat ttgacattca gtgtcaaaag cagcagaggc 1740
tcttctgacc cccaaggggt gacgtgcgga gctgctacac tctctgcaga gagagtcaga 1800
ggggacaaca aggagtatga gtactcagtg gagtgccagg aggacagtgc ctgcccagct 1860
gctgaggaga gtctgcccat tgaggtcatg gtggatgccg ttcacaagct caagtatgaa 1920
aactacacca gcagcttctt catcagggac atcatcaaac ctgacccacc caagaacttg 1980
cagctgaagc cattaaagaa ttctcggcag gtggaggtca gctgggagta ccctgacacc 2040
tggagtactc cacattccta cttctccctg acattctgcg ttcaggtcca gggcaagagc 2100
aagagagaaa agaaagatag agtcttcacg gacaagacct cagccacggt catctgccgc 2160
aaaaatgcca gcattagcgt gcgggcccag gaccgctact atagctcatc ttggagcgaa 2220
tgggcatctg tgccctgcag tggaggcggt ggaagtggcg gtggaggctc tggaggtggc 2280
ggaagcaccc aggactgctc cttccaacac agccccatct cctccgactt cgctgtcaaa 2340
atccgtgagc tgtctgacta cctgcttcaa gattacccag tcaccgtggc ctccaacctg 2400
caggacgagg agctctgcgg gggcctctgg cggctggtcc tggcacagcg ctggatggag 2460
cggctcaaga ctgtcgctgg gtccaagatg caaggcttgc tggagcgcgt gaacacggag 2520
atacactttg tcaccaaatg tgcctttcag ccccccccca gctgtcttcg cttcgtccag 2580
accaacatct cccgcctcct gcaggagacc tccgagcagc tggtggcgct gaagccctgg 2640
atcactcgcc agaacttctc ccggtgcctg gagctgcagt gtcagcccga ctcctcaacc 2700
ctgccacccc catggagtcc ccggcccctg gaggccacag ccccgacagc cccgcagccc 2760
cctctgctcc tcctactgct gctgcccgtg ggcctcctgc tgctggccgc tgcctggtgc 2820
ctgcactggc agaggacgcg gcggaggaca ccccgccctg gggagcaggt gccccccgtc 2880
cccagtcccc aggacctgct gcttgtggag cactga 2916
<210> 55
<211> 2973
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL7IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 55
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagcgattgt gatattgaag gtaaagatgg caaacaatat 840
gagagtgttc taatggtcag catcgatcaa ttattggaca gcatgaaaga aattggtagc 900
aattgcctga ataatgaatt taactttttt aaaagacata tctgtgatgc taataaggaa 960
ggtatgtttt tattccgtgc tgctcgcaag ttgaggcaat ttcttaaaat gaatagcact 1020
ggtgattttg atctccactt attaaaagtt tcagaaggca caacaatact gttgaactgc 1080
actggccagg ttaaaggaag aaaaccagct gccctgggtg aagcccaacc aacaaagagt 1140
ttggaagaaa ataaatcttt aaaggaacag aaaaaactga atgacttgtg tttcctaaag 1200
agactattac aagagataaa aacttgttgg aataaaattt tgatgggcac taaagaacac 1260
gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctatgtgt 1320
caccagcagt tggtcatctc ttggttttcc ctggtttttc tggcatctcc cctcgtggcc 1380
atatgggaac tgaagaaaga tgtttatgtc gtagaattgg attggtatcc ggatgcccct 1440
ggagaaatgg tggtcctcac ctgtgacacc cctgaagaag atggtatcac ctggaccttg 1500
gaccagagca gtgaggtctt aggctctggc aaaaccctga ccatccaagt caaagagttt 1560
ggagatgctg gccagtacac ctgtcacaaa ggaggcgagg ttctaagcca ttcgctcctg 1620
ctgcttcaca aaaaggaaga tggaatttgg tccactgata ttttaaagga ccagaaagaa 1680
cccaaaaata agacctttct aagatgcgag gccaagaatt attctggacg tttcacctgc 1740
tggtggctga cgacaatcag tactgatttg acattcagtg tcaaaagcag cagaggctct 1800
tctgaccccc aaggggtgac gtgcggagct gctacactct ctgcagagag agtcagaggg 1860
gacaacaagg agtatgagta ctcagtggag tgccaggagg acagtgcctg cccagctgct 1920
gaggagagtc tgcccattga ggtcatggtg gatgccgttc acaagctcaa gtatgaaaac 1980
tacaccagca gcttcttcat cagggacatc atcaaacctg acccacccaa gaacttgcag 2040
ctgaagccat taaagaattc tcggcaggtg gaggtcagct gggagtaccc tgacacctgg 2100
agtactccac attcctactt ctccctgaca ttctgcgttc aggtccaggg caagagcaag 2160
agagaaaaga aagatagagt cttcacggac aagacctcag ccacggtcat ctgccgcaaa 2220
aatgccagca ttagcgtgcg ggcccaggac cgctactata gctcatcttg gagcgaatgg 2280
gcatctgtgc cctgcagtgg aggcggtgga agtggcggtg gaggctctgg aggtggcgga 2340
agcacccagg actgctcctt ccaacacagc cccatctcct ccgacttcgc tgtcaaaatc 2400
cgtgagctgt ctgactacct gcttcaagat tacccagtca ccgtggcctc caacctgcag 2460
gacgaggagc tctgcggggg cctctggcgg ctggtcctgg cacagcgctg gatggagcgg 2520
ctcaagactg tcgctgggtc caagatgcaa ggcttgctgg agcgcgtgaa cacggagata 2580
cactttgtca ccaaatgtgc ctttcagccc ccccccagct gtcttcgctt cgtccagacc 2640
aacatctccc gcctcctgca ggagacctcc gagcagctgg tggcgctgaa gccctggatc 2700
actcgccaga acttctcccg gtgcctggag ctgcagtgtc agcccgactc ctcaaccctg 2760
ccacccccat ggagtccccg gcccctggag gccacagccc cgacagcccc gcagccccct 2820
ctgctcctcc tactgctgct gcccgtgggc ctcctgctgc tggccgctgc ctggtgcctg 2880
cactggcaga ggacgcggcg gaggacaccc cgccctgggg agcaggtgcc ccccgtcccc 2940
agtccccagg acctgctgct tgtggagcac tga 2973
<210> 56
<211> 2916
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL15IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 56
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagcggcatt catgtcttca ttttgggctg tttcagtgca 840
gggcttccta aaacagaagc caactgggtg aatgtaataa gtgatttgaa aaaaattgaa 900
gatcttattc aatctatgca tattgatgct actttatata cggaaagtga tgttcacccc 960
agttgcaaag taacagcaat gaagtgcttt ctcttggagt tacaagttat ttcacttgag 1020
tccggagatg caagtattca tgatacagta gaaaatctga tcatcctagc aaacaacagt 1080
ttgtcttcta atgggaatgt aacagaatct ggatgcaaag aatgtgagga actggaggaa 1140
aaaaatatta aagaattttt gcagagtttt gtacatattg tccaaatgtt catcaacact 1200
tctgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 1260
tgtcaccagc agttggtcat ctcttggttt tccctggttt ttctggcatc tcccctcgtg 1320
gccatatggg aactgaagaa agatgtttat gtcgtagaat tggattggta tccggatgcc 1380
cctggagaaa tggtggtcct cacctgtgac acccctgaag aagatggtat cacctggacc 1440
ttggaccaga gcagtgaggt cttaggctct ggcaaaaccc tgaccatcca agtcaaagag 1500
tttggagatg ctggccagta cacctgtcac aaaggaggcg aggttctaag ccattcgctc 1560
ctgctgcttc acaaaaagga agatggaatt tggtccactg atattttaaa ggaccagaaa 1620
gaacccaaaa ataagacctt tctaagatgc gaggccaaga attattctgg acgtttcacc 1680
tgctggtggc tgacgacaat cagtactgat ttgacattca gtgtcaaaag cagcagaggc 1740
tcttctgacc cccaaggggt gacgtgcgga gctgctacac tctctgcaga gagagtcaga 1800
ggggacaaca aggagtatga gtactcagtg gagtgccagg aggacagtgc ctgcccagct 1860
gctgaggaga gtctgcccat tgaggtcatg gtggatgccg ttcacaagct caagtatgaa 1920
aactacacca gcagcttctt catcagggac atcatcaaac ctgacccacc caagaacttg 1980
cagctgaagc cattaaagaa ttctcggcag gtggaggtca gctgggagta ccctgacacc 2040
tggagtactc cacattccta cttctccctg acattctgcg ttcaggtcca gggcaagagc 2100
aagagagaaa agaaagatag agtcttcacg gacaagacct cagccacggt catctgccgc 2160
aaaaatgcca gcattagcgt gcgggcccag gaccgctact atagctcatc ttggagcgaa 2220
tgggcatctg tgccctgcag tggaggcggt ggaagtggcg gtggaggctc tggaggtggc 2280
ggaagcaccc aggactgctc cttccaacac agccccatct cctccgactt cgctgtcaaa 2340
atccgtgagc tgtctgacta cctgcttcaa gattacccag tcaccgtggc ctccaacctg 2400
caggacgagg agctctgcgg gggcctctgg cggctggtcc tggcacagcg ctggatggag 2460
cggctcaaga ctgtcgctgg gtccaagatg caaggcttgc tggagcgcgt gaacacggag 2520
atacactttg tcaccaaatg tgcctttcag ccccccccca gctgtcttcg cttcgtccag 2580
accaacatct cccgcctcct gcaggagacc tccgagcagc tggtggcgct gaagccctgg 2640
atcactcgcc agaacttctc ccggtgcctg gagctgcagt gtcagcccga ctcctcaacc 2700
ctgccacccc catggagtcc ccggcccctg gaggccacag ccccgacagc cccgcagccc 2760
cctctgctcc tcctactgct gctgcccgtg ggcctcctgc tgctggccgc tgcctggtgc 2820
ctgcactggc agaggacgcg gcggaggaca ccccgccctg gggagcaggt gccccccgtc 2880
cccagtcccc aggacctgct gcttgtggag cactga 2916
<210> 57
<211> 2916
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码hIL12aIL21IL12bFLT3L蛋白质异二聚体的核苷酸序列
<400> 57
atgtggcccc ctgggtcagc ctcccagcca ccgccctcac ctgccgcggc cacaggtctg 60
catccagcgg ctcgccctgt gtccctgcag tgccggctca gcatgtgtcc agcgcgcagc 120
ctcctccttg tggctaccct ggtcctcctg gaccacctca gtttggccag aaacctcccc 180
gtggccactc cagacccagg aatgttccca tgccttcacc actcccaaaa cctgctgagg 240
gccgtcagca acatgctcca gaaggccaga caaactctag aattttaccc ttgcacttct 300
gaagagattg atcatgaaga tatcacaaaa gataaaacca gcacagtgga ggcctgttta 360
ccattggaat taaccaagaa tgagagttgc ctaaattcca gagagacctc tttcataact 420
aatgggagtt gcctggcctc cagaaagacc tcttttatga tggccctgtg ccttagtagt 480
atttatgaag acttgaagat gtaccaggtg gagttcaaga ccatgaatgc aaagcttctg 540
atggatccta agaggcagat ctttctagat caaaacatgc tggcagttat tgatgagctg 600
atgcaggccc tgaatttcaa cagtgagact gtgccacaaa aatcctccct tgaagaaccg 660
gatttttata aaactaaaat caagctctgc atacttcttc atgctttcag aattcgggca 720
gtgactattg atagagtgat gagctatctg aatgcttccg gtggaggagg ttctggaggc 780
ggtggaagtg gtggcggagg tagccaaggt caagatcgcc acatgattag aatgcgtcaa 840
cttatagata ttgttgatca gctgaaaaat tatgtgaatg acttggtccc tgaatttctg 900
ccagctccag aagatgtaga gacaaactgt gagtggtcag ctttttcctg ctttcagaag 960
gcccaactaa agtcagcaaa tacaggaaac aatgaaagga taatcaatgt atcaattaaa 1020
aagctgaaga ggaaaccacc ttccacaaat gcagggagaa gacagaaaca cagactaaca 1080
tgcccttcat gtgattctta tgagaaaaaa ccacccaaag aattcctaga aagattcaaa 1140
tcacttctcc aaaagatgat tcatcagcat ctgtcctcta gaacacacgg aagtgaagat 1200
tccgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg 1260
tgtcaccagc agttggtcat ctcttggttt tccctggttt ttctggcatc tcccctcgtg 1320
gccatatggg aactgaagaa agatgtttat gtcgtagaat tggattggta tccggatgcc 1380
cctggagaaa tggtggtcct cacctgtgac acccctgaag aagatggtat cacctggacc 1440
ttggaccaga gcagtgaggt cttaggctct ggcaaaaccc tgaccatcca agtcaaagag 1500
tttggagatg ctggccagta cacctgtcac aaaggaggcg aggttctaag ccattcgctc 1560
ctgctgcttc acaaaaagga agatggaatt tggtccactg atattttaaa ggaccagaaa 1620
gaacccaaaa ataagacctt tctaagatgc gaggccaaga attattctgg acgtttcacc 1680
tgctggtggc tgacgacaat cagtactgat ttgacattca gtgtcaaaag cagcagaggc 1740
tcttctgacc cccaaggggt gacgtgcgga gctgctacac tctctgcaga gagagtcaga 1800
ggggacaaca aggagtatga gtactcagtg gagtgccagg aggacagtgc ctgcccagct 1860
gctgaggaga gtctgcccat tgaggtcatg gtggatgccg ttcacaagct caagtatgaa 1920
aactacacca gcagcttctt catcagggac atcatcaaac ctgacccacc caagaacttg 1980
cagctgaagc cattaaagaa ttctcggcag gtggaggtca gctgggagta ccctgacacc 2040
tggagtactc cacattccta cttctccctg acattctgcg ttcaggtcca gggcaagagc 2100
aagagagaaa agaaagatag agtcttcacg gacaagacct cagccacggt catctgccgc 2160
aaaaatgcca gcattagcgt gcgggcccag gaccgctact atagctcatc ttggagcgaa 2220
tgggcatctg tgccctgcag tggaggcggt ggaagtggcg gtggaggctc tggaggtggc 2280
ggaagcaccc aggactgctc cttccaacac agccccatct cctccgactt cgctgtcaaa 2340
atccgtgagc tgtctgacta cctgcttcaa gattacccag tcaccgtggc ctccaacctg 2400
caggacgagg agctctgcgg gggcctctgg cggctggtcc tggcacagcg ctggatggag 2460
cggctcaaga ctgtcgctgg gtccaagatg caaggcttgc tggagcgcgt gaacacggag 2520
atacactttg tcaccaaatg tgcctttcag ccccccccca gctgtcttcg cttcgtccag 2580
accaacatct cccgcctcct gcaggagacc tccgagcagc tggtggcgct gaagccctgg 2640
atcactcgcc agaacttctc ccggtgcctg gagctgcagt gtcagcccga ctcctcaacc 2700
ctgccacccc catggagtcc ccggcccctg gaggccacag ccccgacagc cccgcagccc 2760
cctctgctcc tcctactgct gctgcccgtg ggcctcctgc tgctggccgc tgcctggtgc 2820
ctgcactggc agaggacgcg gcggaggaca ccccgccctg gggagcaggt gccccccgtc 2880
cccagtcccc aggacctgct gcttgtggag cactga 2916
<210> 58
<211> 18
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 2A自切割肽氨基酸序列
<400> 58
Glu Gly Arg Gly Ser Leu Leu Thr Cys Gly Asp Val Glu Glu Asn Pro
1 5 10 15
Gly Pro
<210> 59
<211> 54
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 编码2A自切割肽的核苷酸序列
<400> 59
gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccct 54
<210> 60
<211> 357
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12a-mIL2第一多肽链
<400> 60
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
210 215 220
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
225 230 235 240
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
245 250 255
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
260 265 270
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
275 280 285
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
290 295 300
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
305 310 315 320
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
325 330 335
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
340 345 350
Ser Thr Ser Pro Gln
355
<210> 61
<211> 357
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL2-mIL12a第一多肽链
<400> 61
Ala Pro Thr Ser Ser Ser Thr Ser Ser Ser Thr Ala Glu Ala Gln Gln
1 5 10 15
Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Leu Glu Gln Leu Leu
20 25 30
Met Asp Leu Gln Glu Leu Leu Ser Arg Met Glu Asn Tyr Arg Asn Leu
35 40 45
Lys Leu Pro Arg Met Leu Thr Phe Lys Phe Tyr Leu Pro Lys Gln Ala
50 55 60
Thr Glu Leu Lys Asp Leu Gln Cys Leu Glu Asp Glu Leu Gly Pro Leu
65 70 75 80
Arg His Val Leu Asp Leu Thr Gln Ser Lys Ser Phe Gln Leu Glu Asp
85 90 95
Ala Glu Asn Phe Ile Ser Asn Ile Arg Val Thr Val Val Lys Leu Lys
100 105 110
Gly Ser Asp Asn Thr Phe Glu Cys Gln Phe Asp Asp Glu Ser Ala Thr
115 120 125
Val Val Asp Phe Leu Arg Arg Trp Ile Ala Phe Cys Gln Ser Ile Ile
130 135 140
Ser Thr Ser Pro Gln Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
145 150 155 160
Gly Gly Gly Ser Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu
165 170 175
Ser Gln Ser Arg Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr
180 185 190
Ala Arg Glu Lys Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp
195 200 205
His Glu Asp Ile Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu
210 215 220
Pro Leu Glu Leu His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr
225 230 235 240
Ser Ser Thr Thr Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu
245 250 255
Met Met Thr Leu Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr
260 265 270
Gln Thr Glu Phe Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His
275 280 285
Gln Gln Ile Ile Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu
290 295 300
Met Gln Ser Leu Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro
305 310 315 320
Val Gly Glu Ala Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu
325 330 335
Leu His Ala Phe Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly
340 345 350
Tyr Leu Ser Ser Ala
355
<210> 62
<211> 332
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12a-mGMCSF第一多肽链
<400> 62
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val
210 215 220
Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr
225 230 235 240
Leu Asn Glu Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys
245 250 255
Leu Thr Cys Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg
260 265 270
Gly Asn Phe Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr
275 280 285
Tyr Gln Thr Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln
290 295 300
Val Thr Thr Tyr Ala Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr
305 310 315 320
Asp Ile Pro Phe Glu Cys Lys Lys Pro Gly Gln Lys
325 330
<210> 63
<211> 337
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12a-mIL7第一多肽链
<400> 63
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Glu Cys His Ile Lys Asp Lys Glu Gly Lys Ala Tyr Glu Ser Val Leu
210 215 220
Met Ile Ser Ile Asp Glu Leu Asp Lys Met Thr Gly Thr Asp Ser Asn
225 230 235 240
Cys Pro Asn Asn Glu Pro Asn Phe Phe Arg Lys His Val Cys Asp Asp
245 250 255
Thr Lys Glu Ala Ala Phe Leu Asn Arg Ala Ala Arg Lys Leu Lys Gln
260 265 270
Phe Leu Lys Met Asn Ile Ser Glu Glu Phe Asn Val His Leu Leu Thr
275 280 285
Val Ser Gln Gly Thr Gln Thr Leu Val Asn Cys Thr Ser Lys Glu Glu
290 295 300
Lys Asn Val Lys Glu Gln Lys Lys Asn Asp Ala Cys Phe Leu Lys Arg
305 310 315 320
Leu Leu Arg Glu Ile Lys Thr Cys Trp Asn Lys Ile Leu Lys Gly Ser
325 330 335
Ile
<210> 64
<211> 341
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12a-mIL15第一多肽链
<400> 64
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
Gly Ile His Val Phe Ile Leu Gly Cys Val Ser Val Gly Leu Pro Lys
210 215 220
Thr Glu Ala Asn Trp Ile Asp Val Arg Tyr Asp Leu Glu Lys Ile Glu
225 230 235 240
Ser Leu Ile Gln Ser Ile His Ile Asp Thr Thr Leu Tyr Thr Asp Ser
245 250 255
Asp Phe His Pro Ser Cys Lys Val Thr Ala Met Asn Cys Phe Leu Leu
260 265 270
Glu Leu Gln Val Ile Leu His Glu Tyr Ser Asn Met Thr Leu Asn Glu
275 280 285
Thr Val Arg Asn Val Leu Tyr Leu Ala Asn Ser Thr Leu Ser Ser Asn
290 295 300
Lys Asn Val Ala Glu Ser Gly Cys Lys Glu Cys Glu Glu Leu Glu Glu
305 310 315 320
Lys Thr Phe Thr Glu Phe Leu Gln Ser Phe Ile Arg Ile Val Gln Met
325 330 335
Phe Ile Asn Thr Ser
340
<210> 65
<211> 337
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12a-mIL21第一多肽链
<400> 65
Arg Val Ile Pro Val Ser Gly Pro Ala Arg Cys Leu Ser Gln Ser Arg
1 5 10 15
Asn Leu Leu Lys Thr Thr Asp Asp Met Val Lys Thr Ala Arg Glu Lys
20 25 30
Leu Lys His Tyr Ser Cys Thr Ala Glu Asp Ile Asp His Glu Asp Ile
35 40 45
Thr Arg Asp Gln Thr Ser Thr Leu Lys Thr Cys Leu Pro Leu Glu Leu
50 55 60
His Lys Asn Glu Ser Cys Leu Ala Thr Arg Glu Thr Ser Ser Thr Thr
65 70 75 80
Arg Gly Ser Cys Leu Pro Pro Gln Lys Thr Ser Leu Met Met Thr Leu
85 90 95
Cys Leu Gly Ser Ile Tyr Glu Asp Leu Lys Met Tyr Gln Thr Glu Phe
100 105 110
Gln Ala Ile Asn Ala Ala Leu Gln Asn His Asn His Gln Gln Ile Ile
115 120 125
Leu Asp Lys Gly Met Leu Val Ala Ile Asp Glu Leu Met Gln Ser Leu
130 135 140
Asn His Asn Gly Glu Thr Leu Arg Gln Lys Pro Pro Val Gly Glu Ala
145 150 155 160
Asp Pro Tyr Arg Val Lys Met Lys Leu Cys Ile Leu Leu His Ala Phe
165 170 175
Ser Thr Arg Val Val Thr Ile Asn Arg Val Met Gly Tyr Leu Ser Ser
180 185 190
Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
195 200 205
His Lys Ser Ser Pro Gln Gly Pro Asp Arg Leu Leu Ile Arg Leu Arg
210 215 220
His Leu Ile Asp Ile Val Glu Gln Leu Lys Ile Tyr Glu Asn Asp Leu
225 230 235 240
Asp Pro Glu Leu Leu Ser Ala Pro Gln Asp Val Lys Gly His Cys Glu
245 250 255
His Ala Ala Phe Ala Cys Phe Gln Lys Ala Lys Leu Lys Pro Ser Asn
260 265 270
Pro Gly Asn Asn Lys Thr Phe Ile Ile Asp Leu Val Ala Gln Leu Arg
275 280 285
Arg Arg Leu Pro Ala Arg Arg Gly Gly Lys Lys Gln Lys His Ile Ala
290 295 300
Lys Cys Pro Ser Cys Asp Ser Tyr Glu Lys Arg Thr Pro Lys Glu Phe
305 310 315 320
Leu Glu Arg Leu Lys Trp Leu Leu Gln Lys Met Ile His Gln His Leu
325 330 335
Ser
<210> 66
<211> 452
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12b-mGMCSF第二多肽链
<400> 66
Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr
1 5 10 15
Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys
85 90 95
Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser
100 105 110
Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys
115 120 125
Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr
130 135 140
Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg
145 150 155 160
Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro
165 170 175
Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln
180 185 190
Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile
195 200 205
Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn
210 215 220
Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro
225 230 235 240
His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys
245 250 255
Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe
260 265 270
Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val
275 280 285
Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp
290 295 300
Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly
305 310 315 320
Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Arg Ser Pro Ile Thr
325 330 335
Val Thr Arg Pro Trp Lys His Val Glu Ala Ile Lys Glu Ala Leu Asn
340 345 350
Leu Leu Asp Asp Met Pro Val Thr Leu Asn Glu Glu Val Glu Val Val
355 360 365
Ser Asn Glu Phe Ser Phe Lys Lys Leu Thr Cys Val Gln Thr Arg Leu
370 375 380
Lys Ile Phe Glu Gln Gly Leu Arg Gly Asn Phe Thr Lys Leu Lys Gly
385 390 395 400
Ala Leu Asn Met Thr Ala Ser Tyr Tyr Gln Thr Tyr Cys Pro Pro Thr
405 410 415
Pro Glu Thr Asp Cys Glu Thr Gln Val Thr Thr Tyr Ala Asp Phe Ile
420 425 430
Asp Ser Leu Lys Thr Phe Leu Thr Asp Ile Pro Phe Glu Cys Lys Lys
435 440 445
Pro Gly Gln Lys
450
<210> 67
<211> 452
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mGMCSF-mIL12b第二多肽链
<400> 67
Ala Pro Thr Arg Ser Pro Ile Thr Val Thr Arg Pro Trp Lys His Val
1 5 10 15
Glu Ala Ile Lys Glu Ala Leu Asn Leu Leu Asp Asp Met Pro Val Thr
20 25 30
Leu Asn Glu Glu Val Glu Val Val Ser Asn Glu Phe Ser Phe Lys Lys
35 40 45
Leu Thr Cys Val Gln Thr Arg Leu Lys Ile Phe Glu Gln Gly Leu Arg
50 55 60
Gly Asn Phe Thr Lys Leu Lys Gly Ala Leu Asn Met Thr Ala Ser Tyr
65 70 75 80
Tyr Gln Thr Tyr Cys Pro Pro Thr Pro Glu Thr Asp Cys Glu Thr Gln
85 90 95
Val Thr Thr Tyr Ala Asp Phe Ile Asp Ser Leu Lys Thr Phe Leu Thr
100 105 110
Asp Ile Pro Phe Glu Cys Lys Lys Pro Gly Gln Lys Gly Gly Gly Gly
115 120 125
Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Met Trp Glu Leu Glu
130 135 140
Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr Pro Asp Ala Pro Gly
145 150 155 160
Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu Glu Asp Asp Ile Thr
165 170 175
Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly Ser Gly Lys Thr Leu
180 185 190
Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly Gln Tyr Thr Cys His
195 200 205
Lys Gly Gly Glu Thr Leu Ser His Ser His Leu Leu Leu His Lys Lys
210 215 220
Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys Asn Phe Lys Asn Lys
225 230 235 240
Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser Gly Arg Phe Thr Cys
245 250 255
Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys Phe Asn Ile Lys Ser
260 265 270
Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr Cys Gly Met Ala Ser
275 280 285
Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg Asp Tyr Glu Lys Tyr
290 295 300
Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro Thr Ala Glu Glu Thr
305 310 315 320
Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln Gln Asn Lys Tyr Glu
325 330 335
Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile Ile Lys Pro Asp Pro
340 345 350
Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn Ser Gln Val Glu Val
355 360 365
Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro His Ser Tyr Phe Ser
370 375 380
Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys Glu Lys Met Lys Glu
385 390 395 400
Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe Leu Val Glu Lys Thr
405 410 415
Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val Cys Val Gln Ala Gln
420 425 430
Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp Ala Cys Val Pro Cys
435 440 445
Arg Val Arg Ser
450
<210> 68
<211> 477
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12b-mIL2第二多肽链
<400> 68
Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr
1 5 10 15
Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys
85 90 95
Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser
100 105 110
Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys
115 120 125
Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr
130 135 140
Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg
145 150 155 160
Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro
165 170 175
Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln
180 185 190
Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile
195 200 205
Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn
210 215 220
Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro
225 230 235 240
His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys
245 250 255
Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe
260 265 270
Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val
275 280 285
Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp
290 295 300
Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly
305 310 315 320
Gly Gly Ser Gly Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Ser
325 330 335
Ser Ser Thr Ala Glu Ala Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln
340 345 350
Gln Gln His Leu Glu Gln Leu Leu Met Asp Leu Gln Glu Leu Leu Ser
355 360 365
Arg Met Glu Asn Tyr Arg Asn Leu Lys Leu Pro Arg Met Leu Thr Phe
370 375 380
Lys Phe Tyr Leu Pro Lys Gln Ala Thr Glu Leu Lys Asp Leu Gln Cys
385 390 395 400
Leu Glu Asp Glu Leu Gly Pro Leu Arg His Val Leu Asp Leu Thr Gln
405 410 415
Ser Lys Ser Phe Gln Leu Glu Asp Ala Glu Asn Phe Ile Ser Asn Ile
420 425 430
Arg Val Thr Val Val Lys Leu Lys Gly Ser Asp Asn Thr Phe Glu Cys
435 440 445
Gln Phe Asp Asp Glu Ser Ala Thr Val Val Asp Phe Leu Arg Arg Trp
450 455 460
Ile Ala Phe Cys Gln Ser Ile Ile Ser Thr Ser Pro Gln
465 470 475
<210> 69
<211> 534
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> mIL12b-mFLT3L第二多肽链
<400> 69
Met Trp Glu Leu Glu Lys Asp Val Tyr Val Val Glu Val Asp Trp Thr
1 5 10 15
Pro Asp Ala Pro Gly Glu Thr Val Asn Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Asp Ile Thr Trp Thr Ser Asp Gln Arg His Gly Val Ile Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Thr Val Lys Glu Phe Leu Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Thr Leu Ser His Ser His Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asn Gly Ile Trp Ser Thr Glu Ile Leu Lys
85 90 95
Asn Phe Lys Asn Lys Thr Phe Leu Lys Cys Glu Ala Pro Asn Tyr Ser
100 105 110
Gly Arg Phe Thr Cys Ser Trp Leu Val Gln Arg Asn Met Asp Leu Lys
115 120 125
Phe Asn Ile Lys Ser Ser Ser Ser Ser Pro Asp Ser Arg Ala Val Thr
130 135 140
Cys Gly Met Ala Ser Leu Ser Ala Glu Lys Val Thr Leu Asp Gln Arg
145 150 155 160
Asp Tyr Glu Lys Tyr Ser Val Ser Cys Gln Glu Asp Val Thr Cys Pro
165 170 175
Thr Ala Glu Glu Thr Leu Pro Ile Glu Leu Ala Leu Glu Ala Arg Gln
180 185 190
Gln Asn Lys Tyr Glu Asn Tyr Ser Thr Ser Phe Phe Ile Arg Asp Ile
195 200 205
Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Met Lys Pro Leu Lys Asn
210 215 220
Ser Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Ser Trp Ser Thr Pro
225 230 235 240
His Ser Tyr Phe Ser Leu Lys Phe Phe Val Arg Ile Gln Arg Lys Lys
245 250 255
Glu Lys Met Lys Glu Thr Glu Glu Gly Cys Asn Gln Lys Gly Ala Phe
260 265 270
Leu Val Glu Lys Thr Ser Thr Glu Val Gln Cys Lys Gly Gly Asn Val
275 280 285
Cys Val Gln Ala Gln Asp Arg Tyr Tyr Asn Ser Ser Cys Ser Lys Trp
290 295 300
Ala Cys Val Pro Cys Arg Val Arg Ser Gly Gly Gly Gly Ser Gly Gly
305 310 315 320
Gly Gly Ser Gly Gly Gly Gly Ser Gly Thr Pro Asp Cys Tyr Phe Ser
325 330 335
His Ser Pro Ile Ser Ser Asn Phe Lys Val Lys Phe Arg Glu Leu Thr
340 345 350
Asp His Leu Leu Lys Asp Tyr Pro Val Thr Val Ala Val Asn Leu Gln
355 360 365
Asp Glu Lys His Cys Lys Ala Leu Trp Ser Leu Phe Leu Ala Gln Arg
370 375 380
Trp Ile Glu Gln Leu Lys Thr Val Ala Gly Ser Lys Met Gln Thr Leu
385 390 395 400
Leu Glu Asp Val Asn Thr Glu Ile His Phe Val Thr Ser Cys Thr Phe
405 410 415
Gln Pro Leu Pro Glu Cys Leu Arg Phe Val Gln Thr Asn Ile Ser His
420 425 430
Leu Leu Lys Asp Thr Cys Thr Gln Leu Leu Ala Leu Lys Pro Cys Ile
435 440 445
Gly Lys Ala Cys Gln Asn Phe Ser Arg Cys Leu Glu Val Gln Cys Gln
450 455 460
Pro Asp Ser Ser Thr Leu Leu Pro Pro Arg Ser Pro Ile Ala Leu Glu
465 470 475 480
Ala Thr Glu Leu Pro Glu Pro Arg Pro Arg Gln Leu Leu Leu Leu Leu
485 490 495
Leu Leu Leu Leu Pro Leu Thr Leu Val Leu Leu Ala Ala Ala Trp Gly
500 505 510
Leu Arg Trp Gln Arg Ala Arg Arg Arg Gly Glu Leu His Pro Gly Val
515 520 525
Pro Leu Pro Ser His Pro
530
<210> 70
<211> 345
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12a-hIL2第一多肽链
<400> 70
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu
210 215 220
Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile
225 230 235 240
Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe
245 250 255
Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu
260 265 270
Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys
275 280 285
Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile
290 295 300
Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala
305 310 315 320
Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe
325 330 335
Cys Gln Ser Ile Ile Ser Thr Leu Thr
340 345
<210> 71
<211> 364
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12a-hIL7第一多肽链
<400> 71
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Asp Cys Asp Ile Glu Gly Lys Asp Gly Lys Gln Tyr
210 215 220
Glu Ser Val Leu Met Val Ser Ile Asp Gln Leu Leu Asp Ser Met Lys
225 230 235 240
Glu Ile Gly Ser Asn Cys Leu Asn Asn Glu Phe Asn Phe Phe Lys Arg
245 250 255
His Ile Cys Asp Ala Asn Lys Glu Gly Met Phe Leu Phe Arg Ala Ala
260 265 270
Arg Lys Leu Arg Gln Phe Leu Lys Met Asn Ser Thr Gly Asp Phe Asp
275 280 285
Leu His Leu Leu Lys Val Ser Glu Gly Thr Thr Ile Leu Leu Asn Cys
290 295 300
Thr Gly Gln Val Lys Gly Arg Lys Pro Ala Ala Leu Gly Glu Ala Gln
305 310 315 320
Pro Thr Lys Ser Leu Glu Glu Asn Lys Ser Leu Lys Glu Gln Lys Lys
325 330 335
Leu Asn Asp Leu Cys Phe Leu Lys Arg Leu Leu Gln Glu Ile Lys Thr
340 345 350
Cys Trp Asn Lys Ile Leu Met Gly Thr Lys Glu His
355 360
<210> 72
<211> 345
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12a-hIL15第一多肽链
<400> 72
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Gly Ile His Val Phe Ile Leu Gly Cys Phe Ser Ala
210 215 220
Gly Leu Pro Lys Thr Glu Ala Asn Trp Val Asn Val Ile Ser Asp Leu
225 230 235 240
Lys Lys Ile Glu Asp Leu Ile Gln Ser Met His Ile Asp Ala Thr Leu
245 250 255
Tyr Thr Glu Ser Asp Val His Pro Ser Cys Lys Val Thr Ala Met Lys
260 265 270
Cys Phe Leu Leu Glu Leu Gln Val Ile Ser Leu Glu Ser Gly Asp Ala
275 280 285
Ser Ile His Asp Thr Val Glu Asn Leu Ile Ile Leu Ala Asn Asn Ser
290 295 300
Leu Ser Ser Asn Gly Asn Val Thr Glu Ser Gly Cys Lys Glu Cys Glu
305 310 315 320
Glu Leu Glu Glu Lys Asn Ile Lys Glu Phe Leu Gln Ser Phe Val His
325 330 335
Ile Val Gln Met Phe Ile Asn Thr Ser
340 345
<210> 73
<211> 345
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12a-hIL21第一多肽链
<400> 73
Arg Asn Leu Pro Val Ala Thr Pro Asp Pro Gly Met Phe Pro Cys Leu
1 5 10 15
His His Ser Gln Asn Leu Leu Arg Ala Val Ser Asn Met Leu Gln Lys
20 25 30
Ala Arg Gln Thr Leu Glu Phe Tyr Pro Cys Thr Ser Glu Glu Ile Asp
35 40 45
His Glu Asp Ile Thr Lys Asp Lys Thr Ser Thr Val Glu Ala Cys Leu
50 55 60
Pro Leu Glu Leu Thr Lys Asn Glu Ser Cys Leu Asn Ser Arg Glu Thr
65 70 75 80
Ser Phe Ile Thr Asn Gly Ser Cys Leu Ala Ser Arg Lys Thr Ser Phe
85 90 95
Met Met Ala Leu Cys Leu Ser Ser Ile Tyr Glu Asp Leu Lys Met Tyr
100 105 110
Gln Val Glu Phe Lys Thr Met Asn Ala Lys Leu Leu Met Asp Pro Lys
115 120 125
Arg Gln Ile Phe Leu Asp Gln Asn Met Leu Ala Val Ile Asp Glu Leu
130 135 140
Met Gln Ala Leu Asn Phe Asn Ser Glu Thr Val Pro Gln Lys Ser Ser
145 150 155 160
Leu Glu Glu Pro Asp Phe Tyr Lys Thr Lys Ile Lys Leu Cys Ile Leu
165 170 175
Leu His Ala Phe Arg Ile Arg Ala Val Thr Ile Asp Arg Val Met Ser
180 185 190
Tyr Leu Asn Ala Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly
195 200 205
Gly Gly Gly Ser Gln Gly Gln Asp Arg His Met Ile Arg Met Arg Gln
210 215 220
Leu Ile Asp Ile Val Asp Gln Leu Lys Asn Tyr Val Asn Asp Leu Val
225 230 235 240
Pro Glu Phe Leu Pro Ala Pro Glu Asp Val Glu Thr Asn Cys Glu Trp
245 250 255
Ser Ala Phe Ser Cys Phe Gln Lys Ala Gln Leu Lys Ser Ala Asn Thr
260 265 270
Gly Asn Asn Glu Arg Ile Ile Asn Val Ser Ile Lys Lys Leu Lys Arg
275 280 285
Lys Pro Pro Ser Thr Asn Ala Gly Arg Arg Gln Lys His Arg Leu Thr
290 295 300
Cys Pro Ser Cys Asp Ser Tyr Glu Lys Lys Pro Pro Lys Glu Phe Leu
305 310 315 320
Glu Arg Phe Lys Ser Leu Leu Gln Lys Met Ile His Gln His Leu Ser
325 330 335
Ser Arg Thr His Gly Ser Glu Asp Ser
340 345
<210> 74
<211> 448
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12b-hGMCSF第二多肽链
<400> 74
Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr
1 5 10 15
Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys
85 90 95
Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys
100 105 110
Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr
115 120 125
Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln
130 135 140
Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly
145 150 155 160
Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala
165 170 175
Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala
180 185 190
Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg
195 200 205
Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu
210 215 220
Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp
225 230 235 240
Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln
245 250 255
Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr
260 265 270
Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala
275 280 285
Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro
290 295 300
Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
305 310 315 320
Ser Ala Pro Ala Arg Ser Pro Ser Pro Ser Thr Gln Pro Trp Glu His
325 330 335
Val Asn Ala Ile Gln Glu Ala Arg Arg Leu Leu Asn Leu Ser Arg Asp
340 345 350
Thr Ala Ala Glu Met Asn Glu Thr Val Glu Val Ile Ser Glu Met Phe
355 360 365
Asp Leu Gln Glu Pro Thr Cys Leu Gln Thr Arg Leu Glu Leu Tyr Lys
370 375 380
Gln Gly Leu Arg Gly Ser Leu Thr Lys Leu Lys Gly Pro Leu Thr Met
385 390 395 400
Met Ala Ser His Tyr Lys Gln His Cys Pro Pro Thr Pro Glu Thr Ser
405 410 415
Cys Ala Thr Gln Ile Ile Thr Phe Glu Ser Phe Lys Glu Asn Leu Lys
420 425 430
Asp Phe Leu Leu Val Ile Pro Phe Asp Cys Trp Glu Pro Val Gln Glu
435 440 445
<210> 75
<211> 530
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> hIL12b-hFLT3L第二多肽链
<400> 75
Ile Trp Glu Leu Lys Lys Asp Val Tyr Val Val Glu Leu Asp Trp Tyr
1 5 10 15
Pro Asp Ala Pro Gly Glu Met Val Val Leu Thr Cys Asp Thr Pro Glu
20 25 30
Glu Asp Gly Ile Thr Trp Thr Leu Asp Gln Ser Ser Glu Val Leu Gly
35 40 45
Ser Gly Lys Thr Leu Thr Ile Gln Val Lys Glu Phe Gly Asp Ala Gly
50 55 60
Gln Tyr Thr Cys His Lys Gly Gly Glu Val Leu Ser His Ser Leu Leu
65 70 75 80
Leu Leu His Lys Lys Glu Asp Gly Ile Trp Ser Thr Asp Ile Leu Lys
85 90 95
Asp Gln Lys Glu Pro Lys Asn Lys Thr Phe Leu Arg Cys Glu Ala Lys
100 105 110
Asn Tyr Ser Gly Arg Phe Thr Cys Trp Trp Leu Thr Thr Ile Ser Thr
115 120 125
Asp Leu Thr Phe Ser Val Lys Ser Ser Arg Gly Ser Ser Asp Pro Gln
130 135 140
Gly Val Thr Cys Gly Ala Ala Thr Leu Ser Ala Glu Arg Val Arg Gly
145 150 155 160
Asp Asn Lys Glu Tyr Glu Tyr Ser Val Glu Cys Gln Glu Asp Ser Ala
165 170 175
Cys Pro Ala Ala Glu Glu Ser Leu Pro Ile Glu Val Met Val Asp Ala
180 185 190
Val His Lys Leu Lys Tyr Glu Asn Tyr Thr Ser Ser Phe Phe Ile Arg
195 200 205
Asp Ile Ile Lys Pro Asp Pro Pro Lys Asn Leu Gln Leu Lys Pro Leu
210 215 220
Lys Asn Ser Arg Gln Val Glu Val Ser Trp Glu Tyr Pro Asp Thr Trp
225 230 235 240
Ser Thr Pro His Ser Tyr Phe Ser Leu Thr Phe Cys Val Gln Val Gln
245 250 255
Gly Lys Ser Lys Arg Glu Lys Lys Asp Arg Val Phe Thr Asp Lys Thr
260 265 270
Ser Ala Thr Val Ile Cys Arg Lys Asn Ala Ser Ile Ser Val Arg Ala
275 280 285
Gln Asp Arg Tyr Tyr Ser Ser Ser Trp Ser Glu Trp Ala Ser Val Pro
290 295 300
Cys Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly
305 310 315 320
Ser Thr Gln Asp Cys Ser Phe Gln His Ser Pro Ile Ser Ser Asp Phe
325 330 335
Ala Val Lys Ile Arg Glu Leu Ser Asp Tyr Leu Leu Gln Asp Tyr Pro
340 345 350
Val Thr Val Ala Ser Asn Leu Gln Asp Glu Glu Leu Cys Gly Gly Leu
355 360 365
Trp Arg Leu Val Leu Ala Gln Arg Trp Met Glu Arg Leu Lys Thr Val
370 375 380
Ala Gly Ser Lys Met Gln Gly Leu Leu Glu Arg Val Asn Thr Glu Ile
385 390 395 400
His Phe Val Thr Lys Cys Ala Phe Gln Pro Pro Pro Ser Cys Leu Arg
405 410 415
Phe Val Gln Thr Asn Ile Ser Arg Leu Leu Gln Glu Thr Ser Glu Gln
420 425 430
Leu Val Ala Leu Lys Pro Trp Ile Thr Arg Gln Asn Phe Ser Arg Cys
435 440 445
Leu Glu Leu Gln Cys Gln Pro Asp Ser Ser Thr Leu Pro Pro Pro Trp
450 455 460
Ser Pro Arg Pro Leu Glu Ala Thr Ala Pro Thr Ala Pro Gln Pro Pro
465 470 475 480
Leu Leu Leu Leu Leu Leu Leu Pro Val Gly Leu Leu Leu Leu Ala Ala
485 490 495
Ala Trp Cys Leu His Trp Gln Arg Thr Arg Arg Arg Thr Pro Arg Pro
500 505 510
Gly Glu Gln Val Pro Pro Val Pro Ser Pro Gln Asp Leu Leu Leu Val
515 520 525
Glu His
530

Claims (15)

1.蛋白质异二聚体，其包含第一多肽链以及不同于所述第一多肽链的第二多肽链，其中所述第一多肽链包含IL12a以及与所述IL12a融合的第一因子，所述第二多肽链包含IL12b以及与所述IL12b融合的第二因子，且其中所述第一因子及所述第二因子选自下组：
1)第一因子为IL2，且第二因子为GMCSF；
2)第一因子为IL7，且第二因子为GMCSF；
3)第一因子为IL15，且第二因子为GMCSF；
4)第一因子为IL21，且第二因子为GMCSF；
5)第一因子为IL2，且第二因子为FLT3L；
6)第一因子为IL7，且第二因子为FLT3L；
7)第一因子为IL15，且第二因子为FLT3L；
8)第一因子为IL21，且第二因子为FLT3L；
9)第一因子为GMCSF，且第二因子为IL2；
10)第一因子为GMCSF，且第二因子为IL7；
11)第一因子为GMCSF，且第二因子为IL15；
12)第一因子为GMCSF，且第二因子为IL21；
13)第一因子为FLT3L，且第二因子为IL2；
14)第一因子为FLT3L，且第二因子为IL7；
15)第一因子为FLT3L，且第二因子为IL15；以及，
16)第一因子为FLT3L，且第二因子为IL21。
2.根据权利要求1所述的蛋白质异二聚体，其中，在所述第一多肽链中，所述IL12a的C末端与所述第一因子的N末端直接或间接融合；和/或，在所述第一多肽链中，所述IL12a的N末端与所述第一因子的C末端直接或间接融合。
3.根据权利要求1所述的蛋白质异二聚体，其中，在所述第二多肽链中，所述IL12b的C末端与所述第二因子的N末端直接或间接融合；和/或，在所述第二多肽链中，所述IL12b的N末端与所述第二因子的C末端直接或间接融合。
4.根据权利要求1所述的蛋白质异二聚体，其中所述IL12a、所述IL12b、所述第一因子和所述第二因子来源自哺乳动物。
5.根据权利要求4所述的蛋白质异二聚体，其中所述IL12a、所述IL12b、所述第一因子和所述第二因子的来源相同。
6.根据权利要求1-5中任一项所述的蛋白质异二聚体，其中所述第一多肽链包括如SEQID NO.60-65和SEQ ID NO.70-73中任一项所示的序列。
7.根据权利要求1-5中任一项所述的蛋白质异二聚体，其中所述第二多肽链包括如SEQID NO.66-69和SEQ ID NO.74-75中任一项所示的序列。
8.根据权利要求1-5中任一项所述的蛋白质异二聚体，其中所述蛋白质异二聚体包括如SEQ ID NO.18-37中任一项所示的序列。
9.一种或多种分离的核酸分子，其编码根据权利要求1-8中任一项所述的蛋白质异二聚体。
10.根据权利要求9所述的核酸分子，其包括如SEQ ID NO.38-57中任一项所示的序列。
11.表达载体，其包括编码根据权利要求1-8中任一项所述的蛋白质异二聚体的核苷酸序列。
12.宿主细胞，其包括根据权利要求11所述的表达载体。
13.药物组合物，其包括根据权利要求1-8中任一项所述的蛋白质异二聚体、根据权利要求9或10所述的核酸分子、根据权利要求11所述的载体和/或根据权利要求12所述的宿主细胞，以及药学上可接受的载体。
14.制备根据权利要求1-8中任一项所述的蛋白质异二聚体的方法，其包括以下的步骤：在能够表达根据权利要求1-8中任一项所述的蛋白质异二聚体的情况下，培养根据权利要求12所述的宿主细胞。
15.根据权利要求1-8中任一项所述的蛋白质异二聚体和根据权利要求13所述的药物组合物在制备治疗黑色素瘤药物中的应用。
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