CN113943737A - 一种鸡ctgf基因在抑制鸡前脂肪细胞分化的应用 - Google Patents
一种鸡ctgf基因在抑制鸡前脂肪细胞分化的应用 Download PDFInfo
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Abstract
本发明公开一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用。鸡CTGF基因的核苷酸序列如SEQ ID NO.1所示，其编码的氨基酸序列如SEQ ID NO.2所示。本发明通过添加外源性CTGF重组蛋白以及构建内源性pCMV‑CTGF过表达载体的方法，利用油红O技术、Real‑time PCR技术、Western blot技术从鸡脂肪细胞的脂滴沉积能力、转录组水平、蛋白组水平证明CTGF基因能够抑制鸡前脂肪细胞分化为成熟脂肪细胞，因此，本发明提供了一种CTGF基因在调控鸡前脂肪细胞分化过程中的新用途，在鸡遗传育种和动物营养领域具有实用价值。
Description
技术领域
本发明涉及生物技术领域，具体涉及一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用。
背景技术
肉鸡业在世界范围内，尤其在中国等发展中国家具有良好的发展前景。中国是世界上仅次于美国和巴西的第三大鸡肉生产国，根据联合国粮农组织(FAO)统计，2017年我国鸡肉产量为1344万吨，占全球鸡肉总产量的12.32％。在过去几十年中，经过持续的选育，肉鸡的生长速度得到了极大的提高，然而，对生长速度的过度追求，导致肉鸡出现了脂肪过度沉积的现象。肉鸡腹部脂肪沉积的增加，导致许多不良后果，例如饲料效率降低和生殖性能。因此，家禽育种的主要目标之一是控制肉鸡腹部脂肪的过多沉积。
动物体内脂肪的形成包括脂肪细胞数目增加和体积增大两个方面,脂肪组织中脂肪细胞的过度增殖和分化则会造成脂肪形成加剧从而引发机体脂肪的过度蓄积，抑制脂肪细胞分化对防止动物体内脂肪过度沉积具有特别重要的意义。脂肪细胞分化是从前脂肪细胞分化为成熟脂肪细胞的过程。目前国内外对鸡的脂肪细胞分化的研究报道很少，从现有的研究结果来看，鸡与哺乳动物脂肪细胞分化的机制并不完全相同，为了解决实际生产中肉鸡腹脂过度蓄积的问题，必须有针对性地开展肉鸡腹部脂肪细胞分化的研究。
鸡体脂性状是受多基因调控的复杂性状，如何从众多基因中筛选出影响体脂性状的候选基因是解析体脂性状形成的重要难题，因此要借助分子育种手段进行研究。研究表明人和小鼠等哺乳动物的CTGF在细胞外基质的形成、细胞的黏附和迁移、血管形成、细胞的增殖与纤维化等过程中具有重要作用，拥有广泛而重要的临床意义。然而，由于现有技术中尚无关于特异性过表达鸡CTGF基因表达的方法，阻碍了科研工作者对鸡CTGF基因的功能和调控机制研究，导致人类对鸡CTGF基因的功能和调控机制不清楚。
发明内容
基于以上不足之处，本发明的主要目的是提供一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用。
为实现上述目的，本发明提供了如下方案：一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用，所述的鸡CTGF基因的核苷酸序列如SEQ ID NO.1所示。
进一步的应用，提供了其编码的氨基酸序列如SEQ ID NO.2所示。
进一步的应用，包括以下(1)-(3)中的至少一项：
(1)、通过PCR扩增获得鸡CTGF基因CDS区的全长序列；
(2)、构建带有HA标签的鸡CTGF基因的过表达载体；
(3)、表达鸡CTGF基因所编码的编码氨基酸序列。
进一步的，如上所述PCR扩增所使用的特异性引物为：正向引物：如SEQ ID NO.3所示；反向引物：如SEQ ID NO.4所示。
本发明的另一目的在于提供一种过表达鸡CTGF基因质粒，由鸡CTGF基因CDS序列与pCMV-HA真核表达载体的重组构建而成，核苷酸序列如SEQ ID NO.5所示，氨基酸序列如SEQ ID NO.6所示；其构建方法步骤如下：
(1)、鸡CTGF基因CDS区扩增引物的设计及PCR扩增，
正向引物：如SEQ ID NO.3所示；
反向引物：如SEQ ID NO.4所示；
(2)、CTGF基因DNA片段的回收；
(3)、pCMV-HA真核表达载体的双酶切；
(4)、鸡CTGF基因CDS区PCR扩增产物与线性化pCMV-HA真核表达载体连接及转化；
(5)、CTGF基因质粒DNA的小量提取；
(6)、pCMV-CTGF真核表达载体的鉴定。
本发明的有益效果及优点：本发明通过添加外源性CTGF重组蛋白以及构建内源性pCMV-CTGF过表达载体的方法，利用油红O技术、Real-time PCR技术、Western blot技术从鸡脂肪细胞的脂滴沉积能力、转录组水平、蛋白组水平证明CTGF基因能够抑制鸡前脂肪细胞分化为成熟脂肪细胞，因此，本发明提供了一种CTGF基因在调控鸡前脂肪细胞分化过程中的新用途，在鸡遗传育种和动物营养领域具有实用价值。因此，本发明构建的CTGF过表达载体可以用来制备抑制鸡前脂肪细胞分化的药物，解决实际生产中肉鸡腹脂过度蓄积的问题。
附图说明
图1为不同基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图；其中
(a)CTGF基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图；
(b)为FGF7基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图；
(c)为MYB基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图；
(d)为GJA1基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图；
(e)为LMBR1基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图；
(f)为SLC16A1基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图；
(g)为IGF1R基因在高、低脂系肉鸡腹部脂肪组织中的表达差异检测图。
图2为利用油红O染色技术检测外源性过表达CTGF对鸡前脂肪细胞脂滴沉积影响图。
图3为利用油红O染色技术检测外源性过表达CTGF对鸡前脂肪细胞脂滴沉积的定量响图。
图4为利用Real-timePCR技术检测外源性过表达CTGF对鸡前脂肪细胞分化过程中正向调控因子mRNA的影响图。
图5为利用Real-timePCR技术检测外源性过表达CTGF对鸡前脂肪细胞分化过程中负向调控因子mRNA的影响图。
图6为利用Western Blot技术检测外源性过表达CTGF对鸡前脂肪细胞分化过程中正向调控因子蛋白的影响图。
图7为利用Western Blot技术检测外源性过表达CTGF对鸡前脂肪细胞分化过程中正向调控因子蛋白的定量图。
图8为菌液PCR鉴定CTGF真核表达载体构建结果图。
图9为测序鉴定CTGF真核表达载体构建结果图。
图10为利用Western blot技术对pCMV-CTGF过表达载体的过表达效果检测图。
图11为利用Western blot技术对pCMV-CTGF过表达载体的过表达效果的定量图。
图12为利用油红O染色技术检测内源性过表达CTGF对鸡前脂肪细胞脂滴沉积影响图。
图13为利用油红O染色技术检测内源性过表达CTGF对鸡前脂肪细胞脂滴沉积的定量响图。
图14为利用Real-timePCR技术检测内源性过表达CTGF对鸡前脂肪细胞分化过程中正向调控因子mRNA的影响图。
图15为利用Real-timePCR技术检测内源性过表达CTGF对鸡前脂肪细胞分化过程中负向调控因子mRNA的影响图。
图16为利用Western Blot技术检测内源性过表达CTGF对鸡前脂肪细胞分化过程中正向调控因子蛋白的影响图。
图17为利用Western Blot技术检测内源性过表达CTGF对鸡前脂肪细胞分化过程中正向调控因子蛋白的定量图。
图18为pCMV-CTGF过表达载体的结构示意图。
具体实施方式
本发明利用鸡的60K SNP芯片技术发现了9个可能影响肉鸡腹脂沉积的重要候选基因：SHH、MC2R、CTGF、FGF7、MYB、GJA1、LMBR1、SLC16A1、IGF1R。本发明利用Primer Premier5.0设计了这9个基因的Real-time PCR引物序列，并交给博仕生物公司和英俊生物公司进行合成，Real-time引物序列如下表4所示。本发明利用Real-time PCR的方法检测这9个候选基因在高、低脂系双向选择系鸡腹部脂肪组织中的表达差异，结果发现CTGF基因在低脂系鸡腹部脂肪中的mRNA表达量显著高于高脂系鸡腹部脂肪中的mRNA表达量(P<0.05，图1a)，其中，SHH与MC2R基因因表达量过低没有检测出结果。因此，本发明将CTGF基因作为影响肉鸡腹脂沉积的重要候选基因进行研究。下面对本发明结合附图举例做进一步的说明，下述实施例中的试验方法如无特殊说明，均为常规方法。下述实施例中所用的试验材料，无特殊说明，均为常规生化试剂公司购买得到。
实施例1：细胞培养
永生化的鸡前脂肪细胞系(ICP2)保存在我们的实验室中。将细胞在补充有10％胎牛血清(Gibco)的DMEM/F12培养基(Gibco，Grand Island，NY，美国)中培养，并保持在37℃，5％湿润的CO 2气氛中。
实施例2：总RNA提取及Real-time PCR
(1)总RNA提取
1)取出12孔板，弃掉培养基，用PBS洗3遍。
2)在六孔板的每孔加1mL的Trizol，摇匀，置于冰上静置5min，在震荡板上震15min。
3)吹打细胞，将培养板中的Trizol移入DEPC处理过的1.5mL EP管中，震荡15s。
4)每1mL的Trizol中加入0.2mL的氯仿，用力震荡15s，室温静置5min。
5)4℃，12000rpm，离心15min。
6)将上层水相转移到新的DEPC处理过的1.5mL EP管中，加入等体积的异丙醇，上下颠倒混匀，室温静置10min。
7)4℃，12000rpm，离心10min。
8)弃上清，加入1mL现配的75％乙醇，震荡洗涤RNA一次。
9)4℃，7500rpm，离心5min。
10)弃上清，将装有RNA沉淀的1.5mL离心管置于超净台中风机干燥10min。
11)向RNA沉淀中加入20μL DEPC水进行溶解。
12)紫外分光光度计测定RNA的浓度，然后立即用于反转录或-80℃保存。
(2)反转录
按照PrimeScriptTMRT reagent Kitwith gDNA Eraser(TAKARA)试剂盒说明书进行两步法操作。去除基因组DNA反应条件见表1。
表1去除基因组DNA反应体系
去除基因组DNA反应条件：42℃ 2min；置于冰上。RNA反转录反应体系见表2。
表2反转录PCR反应体系
RNA反转录反应条件：37℃，5min；85℃，5min；4℃保存。
(3)Real-time PCR
按照FastStart Universal SYBR Green Master(Rox)试剂盒说明书操作，Real-time PCR反应体系见表3。
表3Real-time PCR反应体系
将上述试剂充分混合，分装到96孔板中(10μL/孔)。反应条件为95℃，10min；40cycles 95℃，15s，60℃，1min，40个循环。
以TBP为内参，采用2–△CT方法计算目的基因的相对表达量，其中△CT＝CT(目的基因)–CT(TBP)。各基因Real-time PCR引物序列如表4：
表4Real-time PCR引物
实施例3：油红O染色、提取比色及蛋白校正
(1)油红O染色、提取比色
1)除去细胞中的培养基，用PBS清洗3次。每孔加入1mL 4％多聚甲醛固定液(6孔板)，4℃固定30min；此时配制油红O工作液，即油红O原液和去离子水按照3:2的比例混合均匀后，用滤纸过滤(现用现配)。
2)弃去固定液，PBS温和洗涤3次，放入65℃烘箱中烘干。
3)油红O染色工作液避光染色15min。
4)弃去油红O染色液，PBS温和洗涤3次，每孔加入1mL 60％异丙醇，立即弃掉，PBS洗涤3次。
5)每孔加入2mL PBS，倒置显微镜下拍照、存图。
6)弃去PBS，每孔加入1mL 100％异丙醇，用于溶解被染细胞里的油红O，置于水平摇床上晃动15min。
7)用酶标仪在510nm处测定OD值。
(2)蛋白校正
1)取出细胞培养板，弃培养基。
2)用PBS洗细胞2次。
3)12孔板每孔加300μl胰酶消化液，37℃1min，至细胞状态呈流沙状。
4)加入1ml完全培养液中和消化液，反应体系全部移至DEPC处理后的1.5mL离心管中。
5)2000rpm，离心7min，弃去上清。
6)加入1mL PBS，轻轻吹打，重悬细胞。
7)4000rpm，离心10min，弃去上清，-80℃保存。
8)取出冻存的细胞，冰浴助融，加入适量的含有1mM PMSF的细胞裂解液充分震荡直至没有细胞团块，冰上静置30min。
9)12000rpm离心5min，取上清。
10)取少量上清，用BCA方法测定蛋白浓度。
11)用油红O提取比色所得OD值/对应蛋白量，即得到油红O提取比色的校正结果。
实施例4：总蛋白的提取及Western blot
(1)总蛋白的提取
1)以12孔板为例，融解Western及IP细胞裂解液，混匀。取1mL裂解液，在使用前数分钟内加入10μL 100nM的PMSF，使PMSF的最终浓度为1mM。
2)收细胞，加PBS洗3遍，每孔加入75μL的Western及IP细胞裂解液，用100ul移液枪吹打数下，使裂解液与细胞充分接触，置于4°冰箱，裂解30min。
3)充分裂解后，4℃ 12000rpm离心5min，取上清移入新的EP管中，即为细胞的总蛋白。
(2)Western blot
1)采用BCA蛋白浓度测定试剂盒(增强型)测定所收集的蛋白浓度，并将测定的蛋白调整至同等浓度，加入适量的5X SDS-PAGE蛋白上样缓冲液，100℃加热煮沸10min，以充分变性蛋白。
2)待蛋白样品冷却到室温后，每个样品上样20μL，按照75V，40min；120V，45min进行常规电泳。
3)电泳结束后，将海绵、转膜滤纸、胶和NC膜按顺序放入转膜槽中，接通电源，按照0.2A的恒定电流进行转膜1h。
4)将NC膜置于含有5％脱脂奶粉的PBST(封闭液)中，室温在摇床上摇2h。
5)用PBST洗去NC膜上的封闭液，并将NC膜孵育在含有一抗的稀释液中，放置4℃冰箱过夜。
6)用PBST洗膜3次，每次10min；然后将膜孵育在含二抗的封闭液中，室温在摇床上摇1h。
7)用PBST洗膜3次，每次10min，然后用BeyoECL Star(特超敏ECL化学发光试剂盒)进行化学发光成像仪检测。
实施例5：CTGF重组蛋白对鸡前脂肪细胞分化的最适浓度的确定
将从PeproTech公司购买的50ug有生物活性的人CTGF重组蛋白(rCTGF)用250uL柠檬酸盐缓冲液稀释成200ug/mL，放置于-20°冰箱备用。将鸡前脂肪细胞接种在12孔板中，并培养直至达70％融合时，然后用rCTGF和油酸同时处理细胞，使rCTGF终浓度分别为|0、10、50、100、200、400ng/mL，48h后收细胞，利用油红O染色及蛋白矫正技术，检测不同浓度的rCTGF对鸡前脂肪细胞分化过程中脂滴沉积的影响，筛选出rCTGF对鸡前脂肪细胞分化的最适作用浓度。结果显示，与对照组(浓度为0ng/mL的rCTGF)相比，浓度为10、50、100、200ng/mL的rCTGF都能够抑制鸡前脂肪细胞分化过程中脂滴沉积(P<0.05或P<0.01)，其中100ng/mL的rCTGF抑制鸡前脂肪细胞分化过程中脂滴沉积效果最明显(P<0.01，图2-3)，可用于后续的过表达试验。
实施例6：外源性过表达CTGF基因对鸡前脂肪细胞分化标志基因mRNA的影响
为了在mRNA水平上研究rCTGF对鸡前脂肪细胞分化过程中分化标志基因表达的影响，待鸡前脂肪细胞汇合度达到70％左右时进行油酸诱导，同时，用浓度为100ng/mL的rCTGF处理细胞，并将柠檬酸盐缓冲液处理的细胞作为对照组。转染48h后收细胞，利用Real-time PCR的方法检测鸡前脂肪细胞分化过程中分化标志基因(KLF2、GATA2是鸡前脂肪细胞分化负调控因子，PPARr、c/EBPβ、AP2、PLIN是鸡前脂肪细胞分化正调控因子)的表达水平。结果显示，rCTGF组PPARr、AP2、C/EBP/β的表达量显著低于对照组(P<0.05或P<0.01，图4)，rCTGF组CATA2的表达量显著高于对照组(P<0.05，图5)；PLIN、KLF2的表达量无显著变化(图4-5)。
实施例7：外源性过表达CTGF基因对鸡前脂肪细胞分化标志基因蛋白的影响
为了在蛋白水平上研究rCTGF对鸡前脂肪细胞分化过程中分化标志基因表达的影响，待鸡前脂肪细胞汇合度达到70％左右时进行油酸诱导，同时，用浓度为100ng/mL的rCTGF处理细胞，并将柠檬酸盐缓冲液处理的细胞作为对照组。转染48h后收细胞，利用Western blot的方法检测鸡前脂肪细胞分化过程中分化标志基因(PPARr、c/EBPβ)的蛋白表达水平。结果显示，rCTGF组细胞内的CTGF的表达量显著低于对照组(P<0.05)，rCTGF组PPARr、C/EBP/β的表达量显著低于对照组(P<0.05或P<0.01，图6-7)。
实施例8：鸡CTGF基因CDS区扩增引物的设计与合成
根据鸡CTGF基因mRNA序列(NM_204274.1)和ClonExpress II One Step CloningKit试剂盒要求，采用Primer Premier 5.0设计引物扩增CTGF基因的全长CDS区及其上下游20bp碱基序列，其中上游引物(F)和下游引物(R)的5′端和3′端分别引入EcoR I和Xho I酶切位点和15bp同源臂序列(表5中斜体部分序列)。引物由金唯智生物技术有限公司合成，引物信息见表5。
表5鸡CTGF基因CDS克隆引物序列
注：斜体为15bp的同源臂序列，下划线部分为酶切位点序列，上游为内切酶EcoR I位点序列，下游为内切酶Xho I位点序列。
实施例9：鸡CTGF基因CDS区PCR扩增
以鸡前脂肪细胞的mRNA为模板，反转录为cDNA，按照诺唯赞高保真聚合酶(PhantaMaxSuper-fidelity DNA Polymerase)说明书要求。PCR反应体系见表6。
表6鸡CTGF基因CDS区PCR扩增体系
PCR反应条件：95℃预变性3min；95℃变性15s，60℃退火15s，72℃延伸1min，共35个循环；72℃彻底延伸5min，4℃保存，PCR扩增出CTGF基因片段。
实施例10：CTGF基因DNA片段回收
采用1.2％琼脂糖凝胶电泳分离PCR产物，按照Axygen生物技术(杭州)有限公司的DNA凝胶回收试剂盒说明书纯化CTGF的PCR扩增产物：
(1)将含有目的片段条带的琼脂糖凝胶割下，放入1.5mL的离心管中。
(2)每100mg凝胶加入300μL DE-A溶液，置75℃水浴10min，每2min颠倒混匀一次，使凝胶完全溶化。
(3)加入1/2DE-A体积的DE-B溶液，充分混匀。
(4)将混合液移入带有吸附柱的2mL离心管中，12000rpm离心1min，弃掉管中液体。(5)在吸附柱中加入500μL W1液,12000rpm离心30s弃掉管中液体。
(6)在吸附柱中加入700μL W2液，12000rpm离心30s弃掉管中液体。
(7)在吸附柱中加入700μL W2液，12000rpm离心1min弃掉管中液体。
(8)将吸附柱放回收集管中，12,000×g离心1min。
(9)将吸附柱移入一个干净的1.5mL的离心管中，在吸附柱的中央加30μL Eluent缓冲液，静置1min后，12000rpm离心1min，即得CTGF胶回收产物，将所得CTGF基因DNA片段送金唯智公司测序。
实施例11：pCMV-HA载体的双酶切
采用EcoR I和Xho I双酶切pCMV-HA载体，得到线性化质粒。酶切体系见表7。
表7pCMV-HA质粒双酶切体系
将上述反应液在37℃反应2h，产物胶回收后用于下一步的连接。
实施例12：PCR产物与线性化pCMV-HA载体连接
按照Axygen生物技术(杭州)有限公司的ClonExpress II One Step Cloning Kit试剂盒的说明书对CTGF基因PCR产物和线性化的pCMV-HA载体进行连接，
表8连接反应体系
使用移液器轻轻吸打混匀，PCR仪中37℃反应30min。
实施例13：连接产物的转化
按照菌株Trans1-T1 Phage Chemically Comptent Cell的说明书操作：
(1)从-80℃冰箱中取出保存的感受态细胞(100μL)，冰浴助融，将5μL连接产物加入其中，冰浴30min。
(2)42℃水浴热激60s，立即置入冰浴中1～2min，加入1mL LB液体培养基，37℃振荡复苏90min。
(3)4,000rpm离心5min，弃去800μL上清，吹匀后铺于LB平板上，平板含0.1％(V/V)的氨苄青霉素(100mg/mL)，37℃恒温培养12h。
实施例14：CTGF基因质粒DNA的小量提取
按照Axygen生物技术(杭州)有限公司的质粒小提试剂盒说明书操作：
(1)取1-4mL菌液相对比较富集的LB培养基，使用12,000rpm离心1min，将菌体离心到底部，弃上清。
(2)加250μL含有RNase A的Buffer S1将细菌沉淀悬浮起来，并且均匀，不含小菌块。
(3)加250μLBuffer S2，轻轻均匀翻转混合4-6次大概5min，使细菌充分裂解，直至形成透亮的溶液。
(4)加350μL Buffer S3，同样均匀翻转混合6-8次，12,000rpm离心10min。
(5)将裂解的上清液转移到DNA制备管，12,000rpm离心1min，弃滤液。
(6)然后洗涤质粒，分别使用500μL Buffer W1，700μLBuffer W2，12,000rpm离心洗涤，在DNA制备膜正中央加60-80μL Eluent或去离子水，室温静置1min，再12,000rpm离心1min，得到目的质粒。
实施例15：菌液PCR鉴定CTGF真核表达载体是否构建成功
挑取若干个转化的单菌落，接种于20mL LB液体培养基中，放于37℃培养箱，225rpm/min振荡培养12h。将过夜培养的菌液作为扩增模板，以表5中引物序列作为扩增引物，PCR反应体系见表7。将扩增产物进行1.2％的琼脂糖凝胶电泳，如图8结果显示，目的条带大小与理论值相符，表明CTGF真核表达载体构建成功。
实施例16：测序鉴定CTGF真核表达载体是否构建成功
取菌液PCR鉴定正确的菌液200μL用封口膜封好，送金唯智公司测序。取菌液PCR鉴定正确的菌液200μL用封口膜封好，送金唯智公司测序。将测序结果序列与NCBI数据库中鸡CTGF基因mRNA序列(NM_204274.1)中CDS区序列进行比对，如图9结果发现两组序列的所有碱基中只有一个碱基不同，即在CDS区序列的第515个碱基由C变成T，导致脯氨酸(CCG)突变成亮氨酸(CTG)。表明所克隆的片段是CTGF基因目标序列，阳性重组质粒命名为pCMV-CTGF。
实施例17：利用Western blot技术对CTGF真核过表达载体的过表达效果进行检测
为了确认真核表达质粒pCMV-CTGF能正确表达出CTGF蛋白，待鸡细胞长到70％左右的汇合度时进行诱导细胞分化，同时，采用Lipofectamine 3000转染试剂将构建的CTGF真核表达载体pCMV-CTGF和pCMV空质粒，分别转染鸡细胞，转染48、72和96h时，收细胞提取总蛋白，利用Western blot技术检测pCMV-CTGF质粒在鸡前脂肪细胞分化中的表达效果。如图10-11结果显示，pCMV-CTGF在转染48h、72h和96h时，均在细胞内表达出了CTGF蛋白，其中72h和96h的过表达效果最好。
实施例18：内源性过表达CTGF基因对鸡前脂肪细胞脂滴沉积影响
为了在诱导分化48h时获得显著的过表达效果。待ICP2细胞汇合度达到70％左右时分别将pCMV-CTGF真核表达载体(试验组)和pCMV-HA空载体(对照组)转染到鸡前脂肪细胞中，24h后进行油酸诱导细胞分化，分化48h后收细胞进行鸡前脂肪细胞脂滴沉积的检测，油红O染色结果显示，试验组中细胞的脂滴沉积显著高于对照组(P<0.05，图12-13)，表明本发明构建的CTGF过表达载体能够抑制鸡前脂肪细胞的脂滴沉积。
实施例19：内源性过表达CTGF基因对鸡前脂肪细胞分化标志基因mRNA的影响
为了在mRNA水平上研究本发明构建的CTGF过表达载体对鸡前脂肪细胞分化过程中分化标志基因表达的影响，待ICP2细胞汇合度达到70％左右时分别将pCMV-CTGF真核表达载体(试验组)和pCMV-HA空载体(对照组)转染到鸡前脂肪细胞中，24h后进行油酸诱导细胞分化，分化48h后收细胞，利用Real-time PCR的方法检测鸡前脂肪细胞分化过程中分化标志基因(KLF2、GATA2是鸡前脂肪细胞分化负调控因子，PPARr、c/EBPβ、AP2、PLIN是鸡前脂肪细胞分化正调控因子)的表达水平。结果显示，试验组中PPARγ、C/EBP/β的表达量显著低于对照组(P<0.05或P<0.01，图14)，试验组GATA2的表达量显著高于对照组(P<0.05，图15)，PLIN1、KLF2的表达量无显著变化(图14-15)。
实施例20：内源性过表达CTGF基因对鸡前脂肪细胞分化标志基因蛋白的影响
为了在蛋白水平上研究本发明构建的CTGF过表达载体对鸡前脂肪细胞分化过程中分化标志基因表达的影响，待ICP2细胞汇合度达到70％左右时分别将pCMV-CTGF真核表达载体(试验组)和pCMV-HA空载体(对照组)转染到鸡前脂肪细胞中，24h后进行油酸诱导细胞分化，分化48h后收细胞，利用Western blot的方法检测鸡前脂肪细胞分化过程中分化标志基因(PPARr、c/EBPβ)的蛋白表达水平。结果显示，过表达CTGF基因后PPARγ、C/EBP/β的蛋白表达量都有不同程度的降低，其中试验组C/EBP/β的蛋白表达量显著低于对照组(P<0.05，图16-17)。
序列表
<110> 东北农业大学
<120> 一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 2972
<212> DNA
<213> Gallus gallus
<400> 1
cggcagtcac gcagcactct cccgccgggc tcaggagcgg caaagcggag cggagcagcc 60
actgccggcc cagcgcgaac ccggacggca ccgcaccgcg ctgctgaaag acgcctcagc 120
gagaaacgct gcgccccgtc ccgtctcgtc tcacagcgca cagcaacccc aggatgtccc 180
ccgccagcct cgccgtcgcc ctgctgctcg ccctcctcgg cccggtaagc gctctcgccg 240
ctcgccccgc tccctccccg ttgtcccggc ccggcggcgc gactgacggc ctctctcccc 300
tctgtccccc ctgctcagga ggtgcgcggc caggagtgca gcgggcagtg ccagtgcggc 360
tcggggcccg gccccagctg ccccgccggc gtctccctgg tgctcgatgg ctgcggctgc 420
tgccgcgtct gcgccaagca gctgggcgag ctgtgcaccg agcgcgaccc ctgcgaccac 480
cacaaagggc tcttctgcga cttcggctcc cccgccaacc gccgcatcgg cgtctgcacc 540
ggtgagtggg gccgctgggg aagcaggagg cggggagggg gtcccggcgg ccgggacggc 600
gtgtgaccgc ccgcctcctg ccttccctcg cagctcggga cggagcgccc tgcgtcttct 660
ccggcatggt gtaccggagc ggcgagtcct tccagagcag ctgcaagtac cagtgcacct 720
gcctggacgg cgcggtgggc tgcgtgcccc tctgcagcat ggacgtccgc ctgcccagcc 780
ctgactgccc ctaccctcgg cgggtgaagc tccccggcaa gtgctgtgag gagtgggtct 840
gcgacgaggc caaggagcag acggcggtgg gacccgcact ggccggtgag tgggcatccc 900
gcttccagtg gcagcctacg gggaaggggc atctgcgtgg gagctgatgt gggtctgctc 960
cagcagccgg agcccagaat tgagtggggt cgtttctcaa gtggctgcca gatcttctct 1020
gggaggggtg ggagcatcca gcagggtacg atttttaggt ggtgggctca ccgtggttgt 1080
ccgtcccctg cagcctacag actggaagac acttacggcc cagaccccac catgatgcgt 1140
gccaattgcc tggtgcagac tactgagtgg agtgcttgct ccaagacctg tggcatgggc 1200
atctcaacca gggtcaccaa cgataatgct ttctgcagac tggagaagca gagcagactg 1260
tgcatggtca gaccttgcga agctgacctg gaggagaaca tcaaggtaaa gagtctcatc 1320
cttaatgaat gatcttctag gatgcttctg tttcctggca gaaattatta ctgaaaaatg 1380
tagagttctg ttagtaagtt tcaagtgctt gaaatttgag ctttgctcca gaagtgaata 1440
gtctacaact aaatgctcgt aatacttaaa cattatggca ctacataaag caactttgcc 1500
tattgaaaat acaaccagta gtagtgcatc ttcctgctgt taacatgcaa gcttagtgcc 1560
atatgaaagc ataccttcta aacgtgttgt attcacttgc atcctaacag aaaggcaaaa 1620
agtgcattcg caccccaaaa atctccaagc ccatcaagtt tgagctgtct ggctgcacca 1680
gcgtgaagac gtacagagct aagttctgtg gtgtctgcac tgacgggcgc tgctgcacac 1740
cccacagaac agccaccctc cccgtggagt tcaagtgccc cgatggagag atcatgaaaa 1800
ggaaaatgat gttcatcaag acctgtgcct gccactacaa ctgccctgga gacaatgaca 1860
tctttgagtc tctgtactac aggaagatgt atggggacat ggcataaagc cagaaggaga 1920
cgctaactag attctcaact tgaactgatt tgcatctcat tttgtaaaca tgattcaata 1980
gcacaaggta tttaaatcag tttttttttt taatttcaac aaactgctcc atgtgactta 2040
agacaatttg tctactgacc ccaaacagtg gtttgaagaa ggtaaaatgg accgtggaac 2100
cacagcaaga cacagcttca gaacatgctg ttcatgaggt ggtgtgaagg ggttaaggga 2160
aatgcacggg ggaaaagcag actttagtga atgttccctt aatgtggtac agtgttgttt 2220
ttgttttgtt tgtttgtttt ttcatctagt agtgcaattg agaaggagac cataagcatg 2280
tttgctgaca caggtttagt gacagcaaca gctggaatgc aaaccctcac ccagaagagt 2340
gcgaagcaga aggtgttctg tttagtcagg accgtaatgt ttcagctctg acactctgat 2400
tcacatggct tggtcagcaa gaatcagaat cgtgtctatt tgactggaca gcttgcggca 2460
gttaggttac ctgcaacaag ctacttttta ttaatattgt aaatactgtg tatatatatt 2520
tgtacagtta tctaaattaa tttaagtttt gtgcctatgt gtaatgcttt ggtatttcaa 2580
tgatagcctt ctttctttgg aacaagatag gtaggatctg aagcttgtct gacaatgcat 2640
tcaaagtatg aaatggatac tttagtggaa actgttcaga gtgaaatgag gagtggagac 2700
tgagatggag agtgttagac ttgagatgcc tgtatccttg caaagcactc gtctgtctgg 2760
caaaatgcat ctacctccac tttattgatc aagtggcttt aagaaaaatg actcttctgt 2820
agctaatcag tctttccact tgagcatttg tttctttctt tgactatggc tctttttttt 2880
tttttttttg gacagtttat ttgttgagaa gtgtgaccaa aagttacatg tttgcacctt 2940
tttagttgaa aataaagtat ttatattttt ta 2972
<210> 2
<211> 344
<212> PRT
<213> Gallus gallus
<400> 2
Met Ser Pro Ala Ser Leu Ala Val Ala Leu Leu Leu Ala Leu Leu Gly
1 5 10 15
Pro Glu Val Arg Gly Gln Glu Cys Ser Gly Gln Cys Gln Cys Gly Ser
20 25 30
Gly Pro Gly Pro Ser Cys Pro Ala Gly Val Ser Leu Val Leu Asp Gly
35 40 45
Cys Gly Cys Cys Arg Val Cys Ala Lys Gln Leu Gly Glu Leu Cys Thr
50 55 60
Glu Arg Asp Pro Cys Asp His His Lys Gly Leu Phe Cys Asp Phe Gly
65 70 75 80
Ser Pro Ala Asn Arg Arg Ile Gly Val Cys Thr Ala Arg Asp Gly Ala
85 90 95
Pro Cys Val Phe Ser Gly Met Val Tyr Arg Ser Gly Glu Ser Phe Gln
100 105 110
Ser Ser Cys Lys Tyr Gln Cys Thr Cys Leu Asp Gly Ala Val Gly Cys
115 120 125
Val Pro Leu Cys Ser Met Asp Val Arg Leu Pro Ser Pro Asp Cys Pro
130 135 140
Tyr Pro Arg Arg Val Lys Leu Pro Gly Lys Cys Cys Glu Glu Trp Val
145 150 155 160
Cys Asp Glu Ala Lys Glu Gln Thr Ala Val Gly Pro Ala Leu Ala Ala
165 170 175
Tyr Arg Leu Glu Asp Thr Tyr Gly Pro Asp Pro Thr Met Met Arg Ala
180 185 190
Asn Cys Leu Val Gln Thr Thr Glu Trp Ser Ala Cys Ser Lys Thr Cys
195 200 205
Gly Met Gly Ile Ser Thr Arg Val Thr Asn Asp Asn Ala Phe Cys Arg
210 215 220
Leu Glu Lys Gln Ser Arg Leu Cys Met Val Arg Pro Cys Glu Ala Asp
225 230 235 240
Leu Glu Glu Asn Ile Lys Lys Gly Lys Lys Cys Ile Arg Thr Pro Lys
245 250 255
Ile Ser Lys Pro Ile Lys Phe Glu Leu Ser Gly Cys Thr Ser Val Lys
260 265 270
Thr Tyr Arg Ala Lys Phe Cys Gly Val Cys Thr Asp Gly Arg Cys Cys
275 280 285
Thr Pro His Arg Thr Ala Thr Leu Pro Val Glu Phe Lys Cys Pro Asp
290 295 300
Gly Glu Ile Met Lys Arg Lys Met Met Phe Ile Lys Thr Cys Ala Cys
305 310 315 320
His Tyr Asn Cys Pro Gly Asp Asn Asp Ile Phe Glu Ser Leu Tyr Tyr
325 330 335
Arg Lys Met Tyr Gly Asp Met Ala
340
<210> 3
<211> 39
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
tggccatgga ggcccgaatt ccagcgcaca gcaacccca 39
<210> 4
<211> 44
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
ccgcggccgc ggtacctcga ggttagcgtc tccttctggc ttta 44
<210> 5
<211> 4842
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
gagttcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc 60
tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 120
ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 180
gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa 240
tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac 300
atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 360
cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 420
agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 480
ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 540
gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac 600
cgggaccgat ccagcctccg gactctagag gatccggtac tagaggaact gaaaaaccag 660
aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg 720
gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa 780
gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg ggcccaccat gtacccatac 840
gatgttccag attacgctct tatggccatg gaggcccgaa ttccagcgca cagcaacccc 900
aggatgtccc ccgccagcct cgccgtcgcc ctgctgctcg ccctcctcgg cccggaggtg 960
cgcggccagg agtgcagcgg gcagtgccag tgcggctcgg ggcccggccc cagctgcccc 1020
gccggcgtct ccctggtgct cgatggctgc ggctgctgcc gcgtctgcgc caagcagctg 1080
ggcgagctgt gcaccgagcg cgacccctgc gaccaccaca aagggctctt ctgcgacttc 1140
ggctcccccg ccaaccgccg catcggcgtc tgcaccgctc gggacggagc gccctgcgtc 1200
ttctccggca tggtgtaccg gagcggcgag tccttccaga gcagctgcaa gtaccagtgc 1260
acctgcctgg acggcgcggt gggctgcgtg cccctctgca gcatggacgt ccgcctgccc 1320
agccctgact gcccctaccc tcggcgggtg aagctccccg gcaagtgctg tgaggagtgg 1380
gtctgcgacg aggccaagga gcagacggcg gtgggacccg cactggccgc ctacagactg 1440
gaagacactt acggcccaga ccccaccatg atgcgtgcca attgcctggt gcagactact 1500
gagtggagtg cttgctccaa gacctgtggc atgggcatct caaccagggt caccaacgat 1560
aatgctttct gcagactgga gaagcagagc agactgtgca tggtcagacc ttgcgaagct 1620
gacctggagg agaacatcaa gaaaggcaaa aagtgcattc gcaccccaaa aatctccaag 1680
cccatcaagt ttgagctgtc tggctgcacc agcgtgaaga cgtacagagc taagttctgt 1740
ggtgtctgca ctgacgggcg ctgctgcaca ccccacagaa cagccaccct ccccgtggag 1800
ttcaagtgcc ccgatggaga gatcatgaaa aggaaaatga tgttcatcaa gacctgtgcc 1860
tgccactaca actgccctgg agacaatgac atctttgagt ctctgtacta caggaagatg 1920
tatggggaca tggcataaag ccagaaggag acgctaacct cgaggtaccg cggccgcggg 1980
gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga 2040
aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc 2100
tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag 2160
gtgtgggagg ttttttcgga tcctctagag tcgatctgca ggcatgctag cttggcgtaa 2220
tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 2280
cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 2340
attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 2400
tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 2460
ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 2520
gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 2580
ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 2640
cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 2700
ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 2760
accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 2820
catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 2880
gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 2940
tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 3000
agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 3060
actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 3120
gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 3180
aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 3240
gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 3300
aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 3360
atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 3420
gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 3480
atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 3540
ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 3600
cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 3660
agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 3720
cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 3780
tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 3840
agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 3900
gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 3960
gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 4020
ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 4080
tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 4140
tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 4200
gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 4260
caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 4320
atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 4380
gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc 4440
tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag 4500
acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca 4560
gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 4620
agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 4680
aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 4740
tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 4800
ccagggtttt cccagtcacg acgttgtaaa acgacggcca gt 4842
<210> 6
<211> 368
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Leu Met Ala Met Glu Ala Arg
1 5 10 15
Ile Pro Ala His Ser Asn Pro Arg Met Ser Pro Ala Ser Leu Ala Val
20 25 30
Ala Leu Leu Leu Ala Leu Leu Gly Pro Glu Val Arg Gly Gln Glu Cys
35 40 45
Ser Gly Gln Cys Gln Cys Gly Ser Gly Pro Gly Pro Ser Cys Pro Ala
50 55 60
Gly Val Ser Leu Val Leu Asp Gly Cys Gly Cys Cys Arg Val Cys Ala
65 70 75 80
Lys Gln Leu Gly Glu Leu Cys Thr Glu Arg Asp Pro Cys Asp His His
85 90 95
Lys Gly Leu Phe Cys Asp Phe Gly Ser Pro Ala Asn Arg Arg Ile Gly
100 105 110
Val Cys Thr Ala Arg Asp Gly Ala Pro Cys Val Phe Ser Gly Met Val
115 120 125
Tyr Arg Ser Gly Glu Ser Phe Gln Ser Ser Cys Lys Tyr Gln Cys Thr
130 135 140
Cys Leu Asp Gly Ala Val Gly Cys Val Pro Leu Cys Ser Met Asp Val
145 150 155 160
Arg Leu Pro Ser Pro Asp Cys Pro Tyr Pro Arg Arg Val Lys Leu Pro
165 170 175
Gly Lys Cys Cys Glu Glu Trp Val Cys Asp Glu Ala Lys Glu Gln Thr
180 185 190
Ala Val Gly Pro Ala Leu Ala Ala Tyr Arg Leu Glu Asp Thr Tyr Gly
195 200 205
Pro Asp Pro Thr Met Met Arg Ala Asn Cys Leu Val Gln Thr Thr Glu
210 215 220
Trp Ser Ala Cys Ser Lys Thr Cys Gly Met Gly Ile Ser Thr Arg Val
225 230 235 240
Thr Asn Asp Asn Ala Phe Cys Arg Leu Glu Lys Gln Ser Arg Leu Cys
245 250 255
Met Val Arg Pro Cys Glu Ala Asp Leu Glu Glu Asn Ile Lys Lys Gly
260 265 270
Lys Lys Cys Ile Arg Thr Pro Lys Ile Ser Lys Pro Ile Lys Phe Glu
275 280 285
Leu Ser Gly Cys Thr Ser Val Lys Thr Tyr Arg Ala Lys Phe Cys Gly
290 295 300
Val Cys Thr Asp Gly Arg Cys Cys Thr Pro His Arg Thr Ala Thr Leu
305 310 315 320
Pro Val Glu Phe Lys Cys Pro Asp Gly Glu Ile Met Lys Arg Lys Met
325 330 335
Met Phe Ile Lys Thr Cys Ala Cys His Tyr Asn Cys Pro Gly Asp Asn
340 345 350
Asp Ile Phe Glu Ser Leu Tyr Tyr Arg Lys Met Tyr Gly Asp Met Ala
355 360 365
Claims (5)
1.一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用，其特征在于，所述的鸡CTGF基因的核苷酸序列如SEQ ID NO.1所示。
2.根据权利要求1所述的一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用，其特征在于：其编码的氨基酸序列如SEQ ID NO.2所示。
3.根据权利要求1所述的一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用，其特征在于：包括以下(1)-(3)中的至少一项：
(1)、通过PCR扩增获得鸡CTGF基因CDS区的全长序列；
(2)、构建带有HA标签的鸡CTGF基因的过表达载体；
(3)、表达鸡CTGF基因所编码的编码氨基酸序列。
4.根据权利要求3所述的一种鸡CTGF基因在抑制鸡前脂肪细胞分化的应用，其特征在于：所述PCR扩增所使用的特异性引物为：正向引物：如SEQ ID NO.3所示；反向引物：如SEQID NO.4所示。
5.一种过表达鸡CTGF基因质粒，特征在于，由鸡CTGF基因CDS序列与pCMV-HA真核表达载体的重组构建而成；核苷酸序列如SEQ ID NO.5所示，氨基酸序列如SEQ ID NO.6所示；其构建方法步骤如下：
(1)、鸡CTGF基因CDS区扩增引物的设计及PCR扩增，
正向引物：如SEQ ID NO.3所示；
反向引物：如SEQ ID NO.4所示；
(2)、CTGF基因DNA片段的回收；
(3)、pCMV-HA真核表达载体的双酶切；
(4)、鸡CTGF基因CDS区PCR扩增产物与线性化pCMV-HA真核表达载体连接及转化；
(5)、CTGF基因质粒DNA的小量提取；
(6)、pCMV-CTGF真核表达载体的鉴定。
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111500590A (zh) * | 2020-05-11 | 2020-08-07 | 石河子大学 | 一种调控鸡脂肪细胞形成的转录因子gata2的应用 |
| CN114621953A (zh) * | 2022-02-24 | 2022-06-14 | 东北农业大学 | 一种特异性抑制鸡CTGF基因表达的siRNA及其应用 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030113720A1 (en) * | 2000-07-28 | 2003-06-19 | Schebye Xiao Min | cDNAs expressed in adipocyte differentiation |
| CN104278054A (zh) * | 2013-07-04 | 2015-01-14 | 中国农业大学 | 激酶Lats2在调控前体脂肪细胞增殖中的应用 |
| CN114621953A (zh) * | 2022-02-24 | 2022-06-14 | 东北农业大学 | 一种特异性抑制鸡CTGF基因表达的siRNA及其应用 |
-
2021
- 2021-09-22 CN CN202111105809.7A patent/CN113943737A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030113720A1 (en) * | 2000-07-28 | 2003-06-19 | Schebye Xiao Min | cDNAs expressed in adipocyte differentiation |
| CN104278054A (zh) * | 2013-07-04 | 2015-01-14 | 中国农业大学 | 激酶Lats2在调控前体脂肪细胞增殖中的应用 |
| CN114621953A (zh) * | 2022-02-24 | 2022-06-14 | 东北农业大学 | 一种特异性抑制鸡CTGF基因表达的siRNA及其应用 |
Non-Patent Citations (6)
| Title |
|---|
| GENBANK DATABASE: "Gallus gallus isolate bGalGal1 chromosome 3, assembly b GalGal1.pat.whiteleghornlayer.GRCg7w,whole genome shotgun sequence" * |
| ROBERT C . BAXTER 等: "Actions of IGF binding proteins and related proteins in adipose tissue" * |
| TAKASHI NISHIDA 等: "Suppression of adipocyte differentiation by low-intensity pulsed ultrasound via inhibition of insulin signaling and promotion of CCN family protein 2" * |
| 商周春 等: "鸡脂肪细胞分化调控的研究进展" * |
| 张慧 等: "CTGF(CCN2)基因生物学功能及其在家禽上的研究进展" * |
| 杨艳;罗飞宏;: "转化生长因子β超家族介导的脂肪细胞分化新进展" * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111500590A (zh) * | 2020-05-11 | 2020-08-07 | 石河子大学 | 一种调控鸡脂肪细胞形成的转录因子gata2的应用 |
| CN114621953A (zh) * | 2022-02-24 | 2022-06-14 | 东北农业大学 | 一种特异性抑制鸡CTGF基因表达的siRNA及其应用 |
| CN114621953B (zh) * | 2022-02-24 | 2024-04-09 | 东北农业大学 | 一种特异性抑制鸡CTGF基因表达的siRNA及其应用 |
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