CN111172223A - 一种cGAMP生物合成方法 - Google Patents
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Abstract
本发明公开了一种cGAMP生物合成方法。本发明提供了一种合成cGAMP的方法，为用核苷转移酶cGAS和蛋白G3BP1催化反应缓冲液中的ATP、GTP和DNA，收集反应产物，即得到合成的cGAMP；所述反应缓冲液包括ATP、GTP和DNA。本发明的实验证明，蛋白G3BP1能促进核苷转移酶cGAS催化ATP、GTP和DNA合成cGAMP，与已报道的cGAS催化ATP、GTP和DNA合成cGAMP方法相比，能够显著提高cGAMP的合成效率。
Description
技术领域
本发明属于生物技术领域，尤其涉及一种cGAMP生物合成方法。
背景技术
DNA感受器(DNA sensors)是机体识别侵染细胞的病毒DNA并激活免疫反应的关键蛋白质。cGAS是2013年被鉴定到的一个胞内DNA感受器，被认为是DNA介导I型干扰素产生的最主要受体。cGAS作为一种核苷转移酶，催化ATP和GTP形成小分子第二信使cGAMP(CyclicGMP-AMP)，进而启动I型干扰素等炎性细胞因子的表达，从而启动免疫反应。
因此，cGAMP被认为是可以用来调节人体免疫反应的理想药物分子，用于增强人体的抵抗感染能力，以及用于肿瘤的免疫治疗。
cGAMP分子式如下：
已知的cGAMP生物合成方法是：
发明内容
本发明的一个目的是提供一种合成cGAMP的方法。
本发明提供的方法，包括如下步骤：在蛋白G3BP1和DNA的作用下，用核苷转移酶cGAS催化底物ATP和GTP，合成cGAMP。
上述方法中，底物为ATP和GTP；酶为核苷转移酶cGAS；DNA的作用是激活核苷转移酶；蛋白G3BP1的作用是提高酶催化活性。
上述方法中，
所述cGAS、所述G3BP1、所述ATP、所述GTP和所述DNA的配比为8μg：8μg：100nM：100nM：1μg。
上述方法中，本发明使用的DNA与现有技术中用cGAS催化ATP、GTP和DNA反应生成的cGAMP的技术中的DNA相同。本领域技术人员根据现有技术很容易选择相应的DNA。本发明使用的DNA长度为大于45bp的双链DNA(至万bp均可用)。作为本发明的一个实施例，使用的DNA为HT-DNA,大于2000bp本发明采用的是Sigma-Aldrich公司的，货号为D6898)。
上述方法中，
所述底物ATP、GTP和DNA存在于反应缓冲溶液中；
所述反应缓冲液包括ATP、GTP、DNA、MgCl2和HEPES；
所述cGAS、所述G3BP1、所述ATP、所述GTP、所述DNA、所述MgCl2和所述HEPES的配比为8μg：8μg：100nmol：100nmol：1μg：250nmol：1μmol。
上述方法中，
所述反应缓冲液由5mM MgCl2、2mM ATP、2mM GTP、0.02mg/ml DNA、20mM HEPES和水组成；且所述反应缓冲液的pH值为7.5。
上述方法中，
所述在蛋白G3BP1和DNA的作用下，用核苷转移酶cGAS催化底物ATP和GTP，合成cGAMP的方法包括如下步骤：将所述核苷转移酶cGAS和所述蛋白G3BP1溶解在所述反应缓冲液中，反应，得到cGAMP。
上述方法中，所述反应的时间为1小时，所述反应的温度为37度。
上述方法中，在所述催化后，还包括如下步骤：用甲醇和乙腈萃取催化得到的产物，收集上清液，干燥后得到cGAMP。
上述蛋白G3BP1在促进核苷转移酶cGAS催化底物ATP、GTP和DNA合成cGAMP中的应用也是本发明保护的范围。
上述蛋白G3BP1在提高核苷转移酶cGAS催化底物ATP、GTP和DNA合成cGAMP产量中的应用也是本发明保护的范围。
本发明另一个目的是提供一种提高cGAMP产量的方法。
本发明提供的方法，为上述合成cGAMP的方法；所述cGAMP产量高于仅用核苷转移酶cGAS催化ATP、GTP和DNA合成的cGAMP产量。
上述蛋白质cGAS可为人源(序列1)或者鼠源(序列3)或者猪源(序列4)或者cGAS_Bovin(序列5)，各个蛋白均可通过原核表达系统利用HisTrap(GE Healthcare,17-5248-01)纯化；
上述蛋白质G3BP1可为人源(序列1)或者鼠源(序列6)或者G3BP1_Bovin(序列7)，各个蛋白均可通过原核表达系统利用HisTrap(GE Healthcare,17-5248-01)纯化。
本发明中的名词注释如下：
HEPES:4-羟乙基哌嗪乙磺酸
MgCl2:氯化镁
HT-DNA：鲱鱼基因组DNA
ATP：腺嘌呤核苷三磷酸
GTP：鸟嘌呤核苷三磷酸
LC-MS/MS：液相色谱质谱/质谱联用
本发明的实验证明，蛋白G3BP1能促进核苷转移酶cGAS催化ATP、GTP和DNA合成cGAMP，与已报道的cGAS催化ATP、GTP和DNA合成cGAMP方法相比，能够显著提高cGAMP的合成效率。
附图说明
图1为cGAMP产量检测结果。
具体实施方式
下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
下述实施例中蛋白质cGAS可为人源(序列1)或者鼠源(序列3)或者猪源(序列4)或者cGAS_Bovin(序列5)，各个蛋白均可通过原核表达系统利用HisTrap(GE Healthcare,17-5248-01)纯化；
蛋白质G3BP1可为人源(序列2)或者鼠源(序列6)或者G3BP1_Bovin(序列7)，各个蛋白均可通过原核表达系统利用HisTrap(GE Healthcare,17-5248-01)纯化。
下述实施例中反应缓冲液为由5mM MgCl2、2mM ATP、2mM GTP、0.02mg/ml HT-DNA、20mM HEPES和水组成，其pH值为7.5。
下述实施例中缺DNA的反应缓冲液为由5mM MgCl2、2mM ATP、2mM GTP、20mM HEPES和水组成，其pH值为7.5。
实施例1、G3BP1在提高cGAMP产量中的应用
本实施例中的采用的合成方法流程如下：
具体合成方法如下：
1、合成cGAMP
合成cGAMP的方法为如下4组:
ATP+GTP+DNA+cGAS+G3BP1组：将蛋白质cGAS(8μg)与G3BP1(8μg)混合，在反应缓冲液50μL(20mM HEPES,pH 7.5；5mM MgCl2；2mM ATP；2mM GTP；0.02mg/ml HT-DNA)中反应1小时(37℃)，得到含有cGAMP的反应产物；
ATP+GTP+cGAS+G3BP1组：将蛋白质cGAS(8μg)与G3BP1(8μg)混合，在缺DNA的反应缓冲液50μl(20mM HEPES,pH 7.5；5mM MgCl2；2mM ATP；2mM GTP)中反应2小时(37℃)，得到含有cGAMP的反应产物；
ATP+GTP+DNA+G3BP1组：将蛋白质G3BP1(8μg)在反应缓冲液50μl(20mM HEPES,pH7.5；5mM MgCl2；2mM ATP；2mM GTP；0.02mg/ml HT-DNA)中反应2小时(37℃)，得到含有cGAMP的反应产物；
ATP+GTP+DNA+cGAS组：将蛋白质cGAS(8μg)在反应缓冲液50μL(20mM HEPES,pH7.5；5mM MgCl2；2mM ATP；2mM GTP；0.02mg/mL HT-DNA)中反应2小时(37℃)，得到含有cGAMP的反应产物。
2、分离纯化cGAMP
1)分离纯化
向上述50μL含有cGAMP的反应产物中加入100μL甲醇和100μL乙腈，混匀，冰浴10分钟；12000rpm 4℃离心10分钟；吸取上清液置于新的离心管，真空干燥，得到干燥后粉末，即为纯化后cGAMP。
2)LC-MS/MS鉴定
将上述干燥后粉末溶于200μL醋酸铵缓冲液(10mM醋酸铵，0.05％(体积百分含量)醋酸，溶剂是水)；12000rpm离心10分钟(4℃)，取上清并稀释5000倍，通过LC-MS/MS鉴定cGAMP。
检测方法：通过cGAMP标准品(InvivoGen,tlrl-cga23)确定目的产物并生成标准曲线；通过标准曲线计算所测产物中cGAMP含量。
LC-MS/MS鉴定的条件及参数：
液相条件：
色谱仪：ekspert 110
色谱柱：EC 150/2.0NUCLEODUR C18Pyramid,3m
流动相：A相：0.2％甲酸-水溶液
B相：乙腈
梯度洗脱参数：
流速：0.4mL/min；柱温：35℃；进样量：5L
质谱条件：
离子源:电喷雾离子源(ESI)
检测方式:多反应监测(MRM)
离子化方式:正离子扫描
离子源有关参数：
气帘气：20Psi；喷雾电压：5500V；雾化温度：550℃；雾化气：65Psi；辅助气：65Psi；去簇电压：90V；摄入电压：7V；
定量离子对：母离子/子离子：675/524；
定性离子对：母离子/子离子：675/506，
母离子/子离子：675/136。
标准品检测结果如表1所示：
表1
| 标准品浓度(μg/L) | 峰面积 |
| 1 | 3501.79 |
| 5 | 14515.19 |
| 10 | 31834.35 |
| 25 | 73700.04 |
| 50 | 163600.72 |
| 100 | 311451.61 |
标准曲线：y＝3138.62654x-145.65929(r＝0.99947)
样品检测结果如表2所示(每组样品进行三次重复试验)：
表2
cGAMP产量检测结果如图1所示，纵坐标表示每1μg cGAS蛋白催化合成的cGAMP的量(μg)；表明，第四组cGAS与G3BP1混合使用时，cGAMP产量明显强于只含cGAS的第三组。
上述结果表明，G3BP1(GTPase-activatingprotein-(SH3domain)-bindingprotein 1)可以明显提高cGAMP的合成量。
SEQUENCE LISTING
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Leu Phe Ser Arg Asp Leu Ile Asp Lys Pro Ser Lys Glu Phe Leu Ser
465 470 475 480
Lys Gln Ile Glu Tyr Glu Arg Asn Asn Gly Phe Pro Val Phe Trp
485 490 495
<210> 5
<211> 498
<212> PRT
<213> 人工序列
<400> 5
Met Ala Pro Pro Arg Arg Lys Ala Thr Arg Lys Ala Ser Glu Thr Ala
1 5 10 15
Ser Gly Val Ser Ala Pro Cys Val Glu Gly Gly Leu Ser Ala Glu Pro
20 25 30
Ser Glu Pro Ala Ala Val Pro Glu Ala Pro Arg Pro Gly Ala Arg Arg
35 40 45
Cys Gly Ala Ala Gly Ala Ser Gly Ser Arg Arg Glu Lys Ser Arg Leu
50 55 60
Asp Pro Arg Glu Lys Pro Gln Val Arg Ala Arg Ala Ala Arg Ala Glu
65 70 75 80
Asp Gln Ala Glu Gly Pro Ala Ala Pro Thr Ala Asp Ala Glu Pro Pro
85 90 95
Ala Ala Pro Gly His Ser Leu Pro Arg Ala Ser Thr Arg Ser Arg Gly
100 105 110
Thr Ala Ser Ser Ala Arg Ala Arg Arg Pro Gln Ser Gly Pro Pro Glu
115 120 125
Gly Pro Gly Leu Gly Pro Arg Ala Pro Ser Pro His Leu Gly Arg Arg
130 135 140
Glu Glu Ala Pro Gly Ala Trp Lys Pro Arg Ala Val Leu Glu Lys Leu
145 150 155 160
Lys Leu Ser Arg Gln Glu Ile Ser Val Ala Ala Glu Val Val Asn Arg
165 170 175
Leu Gly Asp His Leu Leu Arg Arg Leu Asn Ser Arg Glu Ser Glu Phe
180 185 190
Lys Gly Val Asp Leu Leu Arg Thr Gly Ser Tyr Tyr Glu Arg Val Lys
195 200 205
Ile Ser Ala Pro Asn Glu Phe Asp Leu Met Phe Thr Leu Glu Val Pro
210 215 220
Arg Ile Gln Leu Glu Glu Tyr Cys Asn Ser Ser Ala His Tyr Phe Val
225 230 235 240
Lys Phe Lys Arg Asn Pro Lys Gly Ser Pro Leu Asp Gln Phe Leu Glu
245 250 255
Gly Gly Ile Leu Ser Ala Ser Lys Met Leu Phe Lys Phe Arg Lys Ile
260 265 270
Ile Lys Glu Glu Ile Lys His Ile Glu Asp Thr Asp Val Ile Met Glu
275 280 285
Arg Lys Lys Arg Gly Ser Pro Ala Val Thr Leu Leu Ile Arg Lys Pro
290 295 300
Arg Glu Ile Ser Val Asp Ile Ile Leu Ala Leu Glu Ser Lys Ser Ser
305 310 315 320
Trp Pro Ala Ser Thr Gln Lys Gly Leu Pro Ile Ser Asn Trp Leu Gly
325 330 335
Thr Lys Val Lys Asp Asn Leu Lys Arg Gln Pro Phe Tyr Leu Val Pro
340 345 350
Lys His Ala Lys Glu Gly Ser Leu Phe Gln Glu Glu Thr Trp Arg Leu
355 360 365
Ser Phe Ser His Ile Glu Lys Ala Ile Leu Thr Asn His Gly Gln Thr
370 375 380
Lys Thr Cys Cys Glu Thr Glu Gly Val Lys Cys Cys Arg Lys Glu Cys
385 390 395 400
Leu Lys Leu Met Lys Tyr Leu Leu Glu Gln Leu Lys Lys Lys Phe Gly
405 410 415
Lys Gln Arg Gly Leu Asp Lys Phe Cys Ser Tyr His Val Lys Thr Ala
420 425 430
Phe Leu His Val Cys Thr Gln Asn Pro His Asp Ser Trp Trp Leu Tyr
435 440 445
Lys Asp Leu Glu Leu Cys Phe Asp Asn Cys Val Thr Tyr Phe Leu Gln
450 455 460
Cys Leu Lys Thr Glu His Leu Glu His Tyr Phe Ile Pro Asp Val Leu
465 470 475 480
Ser Lys Gln Ile Glu Tyr Glu Gln Asn Asn Gly Phe Pro Val Phe Asp
485 490 495
Glu Phe
<210> 6
<211> 464
<212> PRT
<213> 人工序列
<400>6
Met Val Met Glu Lys Pro Ser Pro Leu Leu Val Gly Arg Glu Phe Val
1 5 10 15
Arg Gln Tyr Tyr Thr Leu Leu Asn Gln Ala Pro Asp Met Leu His Arg
20 25 30
Phe Tyr Gly Lys Asn Ser Ser Tyr Ala His Gly Gly Leu Asp Ser Asn
35 40 45
Gly Lys Pro Ala Asp Ala Val Tyr Gly Gln Lys Glu Ile His Arg Lys
50 55 60
Val Met Ser Gln Asn Phe Thr Asn Cys His Thr Lys Ile Arg His Val
65 70 75 80
Asp Ala His Ala Thr Leu Asn Asp Gly Val Val Val Gln Val Met Gly
85 90 95
Leu Leu Ser Asn Asn Asn Gln Ala Leu Arg Arg Phe Met Gln Thr Phe
100 105 110
Val Leu Ala Pro Glu Gly Ser Val Ala Asn Lys Phe Tyr Val His Asn
115 120 125
Asp Ile Phe Arg Tyr Gln Asp Glu Val Phe Gly Gly Phe Val Thr Glu
130 135 140
Pro Gln Glu Glu Ser Glu Glu Glu Val Glu Glu Pro Glu Glu Arg Gln
145 150 155 160
Gln Thr Pro Glu Val Val Pro Asp Asp Ser Gly Thr Phe Tyr Asp Gln
165 170 175
Thr Val Ser Asn Asp Leu Glu Glu His Leu Glu Glu Pro Val Val Glu
180 185 190
Pro Glu Pro Glu Pro Glu Pro Glu Pro Glu Pro Glu Pro Val Ser Asp
195 200 205
Ile Gln Glu Asp Lys Pro Glu Ala Ala Leu Glu Glu Ala Ala Pro Asp
210 215 220
Asp Val Gln Lys Ser Thr Ser Pro Ala Pro Ala Asp Val Ala Pro Ala
225 230 235 240
Gln Glu Asp Leu Arg Thr Phe Ser Trp Ala Ser Val Thr Ser Lys Asn
245 250 255
Leu Pro Pro Ser Gly Ala Val Pro Val Thr Gly Thr Pro Pro His Val
260 265 270
Val Lys Val Pro Ala Ser Gln Pro Arg Pro Glu Ser Lys Pro Asp Ser
275 280 285
Gln Ile Pro Pro Gln Arg Pro Gln Arg Asp Gln Arg Val Arg Glu Gln
290 295 300
Arg Ile Asn Ile Pro Pro Gln Arg Gly Pro Arg Pro Ile Arg Glu Ala
305 310 315 320
Gly Glu Pro Gly Asp Val Glu Pro Arg Arg Met Val Arg His Pro Asp
325 330 335
Ser His Gln Leu Phe Ile Gly Asn Leu Pro His Glu Val Asp Lys Ser
340 345 350
Glu Leu Lys Asp Phe Phe Gln Asn Phe Gly Asn Val Val Glu Leu Arg
355 360 365
Ile Asn Ser Gly Gly Lys Leu Pro Asn Phe Gly Phe Val Val Phe Asp
370 375 380
Asp Ser Glu Pro Val Gln Lys Val Leu Ser Asn Arg Pro Ile Met Phe
385 390 395 400
Arg Gly Ala Val Arg Leu Asn Val Glu Glu Lys Lys Thr Arg Ala Ala
405 410 415
Arg Glu Gly Asp Arg Arg Asp Asn Arg Leu Arg Gly Pro Gly Gly Pro
420 425 430
Arg Gly Gly Pro Ser Gly Gly Met Arg Gly Pro Pro Arg Gly Gly Met
435 440 445
Val Gln Lys Pro Gly Phe Gly Val Gly Arg Gly Ile Thr Thr Pro Arg
450 455 460
<210> 7
<211> 465
<212> PRT
<213> 人工序列
<400> 7
Met Val Met Glu Lys Pro Ser Pro Leu Leu Val Gly Arg Glu Phe Val
1 5 10 15
Arg Gln Tyr Tyr Thr Leu Leu Asn Gln Ala Pro Asp Met Leu His Arg
20 25 30
Phe Tyr Gly Lys Asn Ser Ser Tyr Val His Gly Gly Leu Asp Ser Asn
35 40 45
Gly Lys Pro Ala Asp Ala Val Tyr Gly Gln Lys Glu Ile His Arg Lys
50 55 60
Val Met Ser Gln Asn Phe Thr Asn Cys His Thr Lys Ile Arg His Val
65 70 75 80
Asp Ala His Ala Thr Leu Asn Asp Gly Val Val Val Gln Val Met Gly
85 90 95
Leu Leu Ser Asn Asn Asn Gln Ala Leu Arg Arg Phe Met Gln Thr Phe
100 105 110
Val Leu Ala Pro Glu Gly Ser Val Ala Asn Lys Phe Tyr Val His Asn
115 120 125
Asp Ile Phe Arg Tyr Gln Asp Glu Val Phe Gly Gly Phe Ile Thr Glu
130 135 140
Pro Gln Glu Glu Ser Glu Glu Glu Val Glu Glu Pro Glu Glu Arg Gln
145 150 155 160
Gln Thr Pro Glu Val Val Pro Asp Asp Ser Gly Thr Phe Tyr Asp Gln
165 170 175
Thr Val Ser Asn Asp Leu Glu Glu His Leu Glu Glu Pro Val Ala Glu
180 185 190
Pro Glu Pro Glu Pro Glu Pro Glu Pro Glu Gln Glu Pro Val Ser Glu
195 200 205
Val Gln Glu Glu Lys Ser Glu Pro Val Leu Glu Glu Thr Ala Pro Glu
210 215 220
Asp Val Gln Lys Ser Ser Ser Pro Ala Pro Ala Asp Ile Ala Gln Thr
225 230 235 240
Val Gln Glu Asp Leu Arg Thr Phe Ser Trp Ala Ser Val Thr Ser Lys
245 250 255
Asn Leu Pro Pro Ser Gly Ala Val Pro Val Thr Gly Ile Pro Pro His
260 265 270
Val Val Lys Val Pro Ala Ser Gln Pro Arg Pro Glu Ser Lys Pro Glu
275 280 285
Ser Gln Ile Pro Leu Gln Arg Pro Gln Arg Asp Gln Arg Val Arg Glu
290 295 300
Gln Arg Ile Asn Val Pro Pro Gln Arg Gly Pro Arg Pro Val Arg Glu
305 310 315 320
Ala Gly Glu Gln Gly Asp Val Glu Pro Arg Arg Ile Val Arg His Pro
325 330 335
Asp Ser His Gln Leu Phe Ile Gly Asn Leu Pro His Glu Val Asp Lys
340 345 350
Ser Glu Leu Lys Asp Phe Phe Gln Asn Tyr Gly Asn Val Val Glu Leu
355 360 365
Arg Ile Asn Ser Gly Gly Lys Leu Pro Asn Phe Gly Phe Val Val Phe
370 375 380
Asp Asp Ser Glu Pro Val Gln Lys Val Leu Ser Asn Arg Pro Ile Met
385 390 395 400
Phe Arg Gly Glu Val Arg Leu Asn Val Glu Glu Lys Lys Thr Arg Ala
405 410 415
Ala Arg Glu Gly Asp Arg Arg Asp Asn Arg Leu Arg Gly Pro Gly Gly
420 425 430
Pro Arg Gly Gly Leu Gly Gly Gly Met Arg Gly Pro Pro Arg Gly Gly
435 440 445
Met Val Gln Lys Pro Gly Phe Gly Val Gly Arg Ser Ile Ala Pro Arg
450 455 460
Gln
465
Claims (10)
1.一种合成cGAMP的方法，包括如下步骤：在蛋白G3BP1和DNA的作用下，用核苷转移酶cGAS催化底物ATP和GTP，合成cGAMP。
2.根据权利要求1所述的方法，其特征在于：
所述cGAS、所述G3BP1、所述ATP、所述GTP和所述DNA的配比为8μg：8μg：100nM：100nM：1μg。
3.根据权利要求1或2所述的方法，其特征在于：
所述底物ATP、GTP和DNA均存在于反应缓冲溶液中；
所述反应缓冲液包括ATP、GTP、DNA、MgCl2和HEPES；
所述cGAS、所述G3BP1、所述ATP、所述GTP、所述DNA、所述MgCl2和所述HEPES的配比为8μg：8μg：100nmol：100nmol：1μg：250nmol：1μmol。
4.根据权利要求3所述的方法，其特征在于：所述反应缓冲液由5mM MgCl2、2mM ATP、2mMGTP、0.02mg/ml DNA、20mM HEPES和水组成；且所述反应缓冲液的pH值为7.5。
5.根据权利要求3或4所述的方法，其特征在于：
所述在蛋白G3BP1和DNA的作用下，用核苷转移酶cGAS催化底物ATP和GTP，合成cGAMP的方法包括如下步骤：将所述核苷转移酶cGAS和所述蛋白G3BP1溶解在所述反应缓冲液中，反应，得到cGAMP。
6.根据权利要求5所述的方法，其特征在于：所述反应的时间为1小时，所述反应的温度为37度。
7.根据权利要求1-6中任一所述的方法，其特征在于：
在所述催化后，还包括如下步骤：用甲醇和乙腈萃取催化得到的产物，收集上清液，干燥后得到cGAMP。
8.蛋白G3BP1在促进核苷转移酶cGAS催化底物ATP、GTP和DNA合成cGAMP中的应用。
9.蛋白G3BP1在提高核苷转移酶cGAS催化底物ATP、GTP和DNA合成cGAMP产量中的应用。
10.一种提高cGAMP产量的方法，为权利要求1-7任一所述方法合成cGAMP；所述cGAMP产量高于仅用核苷转移酶cGAS催化ATP、GTP和DNA合成的cGAMP产量。
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114796451A (zh) * | 2022-02-09 | 2022-07-29 | 浙江瑞吉康生物医药有限公司 | 使用多肽治疗白内障的方法 |
Citations (1)
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| CN107106589A (zh) * | 2014-12-17 | 2017-08-29 | 立博美华基因科技有限责任公司 | 用cGAMP或cGAsMP治疗癌症的方法 |
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| CN107106589A (zh) * | 2014-12-17 | 2017-08-29 | 立博美华基因科技有限责任公司 | 用cGAMP或cGAsMP治疗癌症的方法 |
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| 刘朝山: "G3BP1通过cGAS调控抗病毒天然免疫的作用机制研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN114796451A (zh) * | 2022-02-09 | 2022-07-29 | 浙江瑞吉康生物医药有限公司 | 使用多肽治疗白内障的方法 |
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