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家蚕蚕蛹变应原的鉴定、分离与纯化 被引量:17
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作者 张杰 刘志刚 +2 位作者 林格 叶湘锋 刘瑞涛 《热带医学杂志》 CAS 2006年第5期496-498,共3页
目的鉴定、分离和纯化大造(Dazao)品系家蚕(Bombyxmori)的蚕蛹粗提液中的变应原蛋白质组分,为家蚕变应原的蛋白质组学和功能基因组学深入研究奠定基础。方法采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和Western-blotting,鉴定了... 目的鉴定、分离和纯化大造(Dazao)品系家蚕(Bombyxmori)的蚕蛹粗提液中的变应原蛋白质组分,为家蚕变应原的蛋白质组学和功能基因组学深入研究奠定基础。方法采用十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和Western-blotting,鉴定了家蚕蚕蛹粗浸液中变应原蛋白质分子,并通过阴离子交换和凝胶过滤层析分别对家蚕蚕蛹变应原蛋白质进行初步纯化。结果在家蚕蚕蛹粗浸液中含有表观分子量分别为80000及30000的蚕蛹变应原;从家蚕蛹全虫粗提液中初步分离到分子量为30000的蛹变应原蛋白组分。结论分离得到的家蚕蚕蛹变应原蛋白组分可用于系统研究家蚕过敏原蛋白分子的蛋白质组学和功能基因组学。 展开更多
关键词 家蚕蚕蛹 过敏原 Western—blotting 离子交换层析
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家蚕蚕蛹过敏原表皮蛋白RR-2基序63前体(CPR63)的克隆表达、纯化鉴定及生物信息学分析 被引量:4
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作者 梁志林 李平 +1 位作者 刘志刚 刘晓宇 《免疫学杂志》 CAS CSCD 北大核心 2016年第7期565-569,共5页
目的克隆表达家蚕蚕蛹表皮蛋白RR-2基序63前体(CPR63)基因,纯化鉴定蛋白的过敏原性,并进行生物信息学分析。方法合成家蚕蚕蛹CPR63蛋白的基因(ref|NM_001173223.1|),将其连接至克隆载体p MD18-T,p MD18-T-CPR63阳性质粒与原核表... 目的克隆表达家蚕蚕蛹表皮蛋白RR-2基序63前体(CPR63)基因,纯化鉴定蛋白的过敏原性,并进行生物信息学分析。方法合成家蚕蚕蛹CPR63蛋白的基因(ref|NM_001173223.1|),将其连接至克隆载体p MD18-T,p MD18-T-CPR63阳性质粒与原核表达载体p ET-44a分别用限制性内切酶酶切,构建出重组表达质粒p ET-44a-CPR63,转化到大肠杆菌BL21(DE3)中经IPTG诱导表达;Western blot检测重组CPR63蛋白特异性过敏原性;用clustalw2和MEGA5工具包进行同源性分析;用Prot Param Tools预测其理化性质;用PSIPRED和SWISS-MODEL预测其蛋白质结构。利用网络服务器IEDB、Preprod和DNAStar,对CPR63蛋白T、B细胞抗原表位进行预测。结果经测序鉴定,本研究成功克隆出了家蚕蚕蛹CPR63基因,其开放阅读框549 bp,编码182组氨基酸。通过大肠杆菌E.coli BL21(DE3)表达CPR63重组蛋白主要以可溶性形式存在,相对分子质量约20 000。Western blot显示重组CPR63能够与家蚕蚕蛹过敏患者血清Ig E结合。家蚕蚕蛹CPR63蛋白与黑脉金斑蝶表皮蛋白RR-2基序63(gb|EHJ69818.1|)同源性为85%。系统进化树结果显示家蚕蚕蛹与小菜蛾亲缘关系比较近。理化性质预测显示CPR63蛋白质较稳定。二级结构及三级结构预测结果显示CPR63的结构主要以无规则卷曲组成。T细胞抗原表位预测得到4个肽序列(5~13、20~28、51~59和87~95)。B细胞抗原表位预测得到4个肽序列(21~40、37~56、47~66和64~83)。结论成功克隆并表达了家蚕蚕蛹CPR63,并证实重组CPR63蛋白具有良好的免疫原性,为进一步研究家蚕蚕蛹过敏原的结构成分及其理化性质奠定理论基础。 展开更多
关键词 家蚕蚕蛹 表皮蛋白RR-2基序63前体(CPR63) 原核表达 免疫原性鉴定 生物信息学分析
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家蚕蚕蛹黄酮类化合物的提取与测定 被引量:1
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作者 张军 宛晓春 +1 位作者 刘朝良 陆翠珍 《安徽农业大学学报》 CAS CSCD 北大核心 2011年第3期386-388,共3页
为推进家蚕蛹体的药用开发,研究家蚕蛹体黄酮类化合物的提取及其含量的测定方法。将蛹体用研钵研碎,选用80%乙醇溶剂在60℃下抽提4 h,1次即可,可有效提取家蚕蛹体黄酮类化合物;以1%AlCl3显色在TU-1800SPC紫外光分光光度计420 nm波长下比... 为推进家蚕蛹体的药用开发,研究家蚕蛹体黄酮类化合物的提取及其含量的测定方法。将蛹体用研钵研碎,选用80%乙醇溶剂在60℃下抽提4 h,1次即可,可有效提取家蚕蛹体黄酮类化合物;以1%AlCl3显色在TU-1800SPC紫外光分光光度计420 nm波长下比色,可测定出家蚕蛹体黄酮类化合物的含量。家蚕蛹体含黄酮类化合物,其质量比达到0.362 mg·g-1左右。 展开更多
关键词 家蚕蚕蛹 黄酮 提取 含量测定
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家蚕蚕蛹基因LOC101743840的原核表达、免疫学鉴定及生物信息学分析 被引量:3
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作者 梁志林 刘晓宇 +1 位作者 刘志刚 陈献雄 《中国热带医学》 CAS 2017年第7期641-645,共5页
目的原核表达家蚕蚕蛹LOC101743840(以下简称LOC)基因,鉴定LOC蛋白的免疫学特性,并进行生物信息学分析。方法将合成的家蚕蚕蛹LOC基因(gi|512917985)连接至克隆载体p MD18-T,用限制性内切酶Xho I和Bam H I分别对p MD18-T-LOC阳性质粒与... 目的原核表达家蚕蚕蛹LOC101743840(以下简称LOC)基因,鉴定LOC蛋白的免疫学特性,并进行生物信息学分析。方法将合成的家蚕蚕蛹LOC基因(gi|512917985)连接至克隆载体p MD18-T,用限制性内切酶Xho I和Bam H I分别对p MD18-T-LOC阳性质粒与原核表达载体p ET-28a双酶切,构建重组表达质粒p ET-28a-LOC并转化到E.coli BL21(DE3)中进行表达(经IPTG诱导);用Western blot鉴定重组LOC蛋白的免疫活性;用Clustalw2和MEGA5工具包分析LOC的基因;用Prot Param Tools预测LOC蛋白的理化性质;用PSIPRED和SWISS-MODEL预测其蛋白质结构。用IEDB、Preprod和DNAStar预测其细胞抗原表位。结果成功表达了家蚕蚕蛹LOC基因,其开放阅读框768 bp,编码255组氨基酸;由E.coli BL21(DE3)原核表达的重组LOC蛋白为可溶性,分子量约33 k D;Western bolt结果显示重组LOC蛋白可特异性结合家蚕蚕蛹过敏患者血清中的特异性Ig E;家蚕蚕蛹LOC与脐橙螟LOC106139919(gi|913330692)蛋白的同源性为69%。系统进化树显示家蚕蚕蛹与美国白蛾亲缘关系比较近。理化性质预测结果显示LOC蛋白质不稳定。蛋白质结构预测结果显示LOC主要的结构为无规则卷曲。T细胞抗原表位预测得到8个肽段(9-17、79-87、93-102、104-115、124-132、152-160、223-231和244-251)。B细胞抗原表位预测得到个5肽段(34-49、59-74、127-142、202-217和213-228)。结论成功表达了家蚕蚕蛹LOC,并证实重组LOC蛋白具有良好的免疫活性,为进一步研究家蚕蚕蛹蛋白的结构成分、免疫学特性及理化性质提供了实验数据。 展开更多
关键词 家蚕蚕蛹 基因LOC101743840 原核表达 免疫学鉴定 生物信息学分析
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Substitution of Silkworm Pupae (Bombyx mori) for Fish Meal in Broodstock Diets for Snakeskin Gourami ( Trichogaster pectoralis) 被引量:1
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作者 Orapint Jintasataporn S. Chumkam Oratal Jintasatapom 《Journal of Agricultural Science and Technology(A)》 2011年第8期1341-1344,共4页
Substitution of silkworm pupae (Bombyx mori) for fish meal in broodstock diets for snakeskin gourami (Trichogaster pectoralis) was conducted in order to evaluate the effect of silkworm pupae on feed digestibility ... Substitution of silkworm pupae (Bombyx mori) for fish meal in broodstock diets for snakeskin gourami (Trichogaster pectoralis) was conducted in order to evaluate the effect of silkworm pupae on feed digestibility and broodstock performance. Snakeskin gourami broodfish were fed with isocaloric diets (DE 3,000 a: 50 Kcal/kg) contained 32% crude protein. The levels of substitution were 0%, 50%, 75% and 100% of protein from fish meal (equal to 0%, 14.57%, 21.75% and 29.03% silk worm by weight). The study was assigned in CRD with 4 treatments and 3 replications. Six hundred of snakeskin gourami with five-month old and average weight of 78.67 ± 2.15 g in female and average weight of 73.00 ± 2.49 g in male were stocked in 12 cages. Fifty fishes of 25 females and 25 males were reared in each cage and fed experimental diet for five months. The result indicated that protein digestibility of test diets were significantly decrease (P 〈 0.05) according to the level of silkworm pupae in diets where as the dry matter digestibility showed no significantly different (P 〉 0.05). The growth performance and egg fecundity were significantly decreased (P 〈 0.05) according to the levels of silkworm pupae. In contrast, egg quality in term of fry number on first spawned, fingering number on first spawned were not significantly different (P 〉 0.05). The survival rate of one month nursing fish on first spawned were significantly different (P 〈 0.05). Hence, silkworm pupae could substitution for 50% of protein from fish meal (14.57% by weight) in snakeskin gourami broodstock diets without any adverse effect on egg quality. 展开更多
关键词 Silkworm pupae fish meal snakeskin gourami broodstock diets
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Production Performance of Broiler Chickens Fed with Silkworm Pupa (Bombyx mon~
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作者 Oratai Jintasataporn 《Journal of Agricultural Science and Technology(A)》 2012年第4期505-510,共6页
The production performance of broilers fed with silkworm pupa (Bombyx mori) was studied by focusing on growth, feed utilization, carcass composition and meat quality. There were two difference sources of silkworm p... The production performance of broilers fed with silkworm pupa (Bombyx mori) was studied by focusing on growth, feed utilization, carcass composition and meat quality. There were two difference sources of silkworm pupa: (1) silkworm pupa from spun silk industry (SSP) and (2) silkworm pupa from silk yam reeling industry (RSP) were mixed into five isonitrogenous (20% CP) and isocaloric (ME = 3,150 kcal/kg feed) diet. The 195 chickens at three weeks old were equally distributed to 15 pens in the closed housing. The experiment was divided into five dietary treatments (three replications each): treatment 1 (10% fishmeal, FM and 0% silkworm pupae) (control)), treatment 2 (0% FM and 10% SSP), treatment 3 (0% FM and 20% SSP), treatment 4 (0% FM and 5% SSP + 5% RSP), and treatment 5 (0% FM and 10% SSP + 10% RSP). Daily weight gain was 0.069, 0.060, 0.051, 0.065 and 0.061 kg/chick in treatments 1, 2, 3, 4 and 5, respectively. It is indicated that replacing fishmeal with silkworm pupa (5%SSP + 5%RSP) performed the same daily weight gain as control but exhibited the higher feed conversion ratio (1.87) than control (1.68). Carcass muscle of control treatment 1 (58.26%) was the highest (P 〈 0.05) and closed to treatment 4 and 5 (57.62% and 57.83%, respectively). Sensory evaluation was not significantly differences (P 〉 0.05). Therefore, treatment 4 (5%SSP + 5%RSP) could be substituted FM with little adverse effect on broiler growth but without any adverse effect on percentage of muscle and sensory test. However, further supplemental silkworm pupa performed reduction in dietary utilization and carcass muscle. 展开更多
关键词 BROILER silkworm pupa production performance CARCASS meat quality
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Productive Performance of Giant Freshwater Prawn (Macrobrachium rosenbergi) Fed Diet Containing Silkworm Pupae (Bombyx mon~ Replacing for Fish Meal
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作者 O. Jintasatapom S. Hatachote C. Thaitungchin 《Journal of Agricultural Science and Technology(A)》 2011年第8期1345-1348,共4页
The productive performance of giant freshwater prawn (Macrobrachium rosenbergii de Man) fed diet containing silkworm pupae (Bombyx mori) replacing for fish meal was conducted by focusing on digestibility, growth p... The productive performance of giant freshwater prawn (Macrobrachium rosenbergii de Man) fed diet containing silkworm pupae (Bombyx mori) replacing for fish meal was conducted by focusing on digestibility, growth performance and feed utilization. Five diets of an isonitrogenous of 35 :e 1% crude protein and isocaloric of digestible energy of 3,200 ±100 kcal/kg were formulated by using silkworm pupae, 0%, 8.6%, 17.2%, 25.8% and 34.7% by weight, replaced for fish meal. Giant freshwater prawn of 19 ± 2.0 g were stocked at 5 ind./m3 in concrete tank of 2.25 ton for three months. The protein digestibility by pepsin enzyme was evaluated in fish meal, silkworm pupae, soybean meal and shrimp meal. The digestibility was significantly difference (P 〈 0.05). The protein digestibility of fishmeal and soybean meal was higher than silkworm pupae and shrimp meal. The productive performance of prawn in terms of growth rate and feed utilization were not significantly different (P 〉 0.05). Therefore, silkworm pupae can be replaced for fishmeal in giant freshwater prawn (Macrobrachium rosenbergii de Man) without any adverse effects on productive performance. 展开更多
关键词 Productive performance giant freshwater prawn silkworm pupae fish meal
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