水貂生长激素的原核表达及特异性抗血清制备

本研究旨在建立水貂生长激素(mink growth hormone,mGH)的原核高效表达体系,探索mGH蛋白规模化生产方法,利用获得的目的蛋白制备特异抗血清。将已构建的原核表达载体pET28b-mGH转化大肠杆菌Rosetta(DE3)TMpLysS感受态细胞,经IPTG诱导表达,超声破碎菌体后离心获得包涵体蛋白,利用8mol/L尿素对其进行溶解和复性,经Ni-NTA树脂亲和层析纯化获得重组蛋白。通过Western blotting检测及MALDI-TOFMS质谱分析对获得的蛋白进行鉴定。用SDS-PAGE电泳法检测蛋白纯度,用BCA法测定其浓度。复性mGH蛋白与完全弗氏佐剂或不完全弗氏佐剂乳化制备免疫抗原,采用皮下多点注射法免疫3只新西兰大白兔,首次免疫剂量为600μg/只,二免剂量为600μg/只,三免剂量为400μg/只。三免10d后分离制备血清,Western blotting法检测其特异性,间接ELISA法检测其效价。结果表明,原核表达的包涵体经Ni-NTA柱纯化后所得蛋白鉴定为mGH融合蛋白;SDS-PAGE电泳鉴定复性mGH蛋白条带单一,纯度达90%以上,BCA法计算蛋白浓度为669μg/mL,均达到了免疫动物的要求;制备的抗血清经Western blotting检测具有良好的结合特异性,间接ELISA法测定其抗体效价达1∶256 000以上。上述结果表明,原核表达载体pET28b-mGH可在大肠杆菌Rosetta(DE3)TMpLysS细胞中高效表达mGH融合蛋白,表达蛋白经变性、复性和纯化后免疫新西兰大白兔,可制备高效价特异性抗血清。     

蛋鸡的支原体病

鸡败血支原体(Mycoplasma gallisepticum,MG)和滑液支原体(Mycoplasma synoviae,MS)在世界许多地区的重型鸡种中已被消灭;然而,在商品蛋鸡中,支原体感染仍为一大问题.最普遍的是 MS 感染,许多鸡群在感染后不表现明显     

Effects of heartworm and intestinal parasitic infections on hematology and peripheral lymph node cytology in louisiana dogs

Hookworm infected dogs had higher blood basophil counts than hookworm negative dogs. Dogs with occult heartworm infection (i.e., without microfilaremia) had higher blood neutrophil counts than heartworm negative dogs. The percentage of eosinophils in peripheral lymph nodes was higher in heartworm infected, microfilaremic dogs than in both occult infected dogs and heartworm negative dogs. Heartworm or intestinal parasitic infection had no other significant effects on hematologic parameters or peripheral lymph node cytology.     

Incarceration of the small intestine by the epiploic foramen in fifteen horses

In 15 horses with acute abdominal disease, a diagnosis of incarceration of small intestine through the epiploic foramen was made, either at the time of exploratory celiotomy or at necropsy.