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用重组鸡抑制素融合蛋白对日本鹌鹑母鸡进行主动免疫可提高生产性能。

Active immunization of Japanese quail hens with a recombinant chicken inhibin fusion protein enhances production performance.

作者信息

Moreau J D, Satterlee D G, Rejman J J, Cadd G G, Kousoulas K G, Fioretti W C

机构信息

Department of Poultry Science, Louisiana Agricultural Experiment Station, Louisiana State University Agricultural Center, Baton Rouge 70803, USA.

出版信息

Poult Sci. 1998 Jun;77(6):894-901. doi: 10.1093/ps/77.6.894.

DOI:10.1093/ps/77.6.894
PMID:9628541
Abstract

The effects of active immunization against inhibin on production performance in female Japanese quail (Coturnix coturnix japonica) were assessed in two separate trials using an MBP-cINA521 fusion protein as an immunogen. The fusion protein, MBP-cINA521, consisted of the bacterial maltose binding protein (MBP) and a truncated form of the mature alpha-subunit of chicken inhibin (cINA521). MBP-cINA1521 was constructed by: 1) excising a 521-bp PstI fragment from a chicken inhibin alpha-subunit cDNA (cINA6; gift of P. A. Johnson), 2) cloning this fragment, which encodes all but the first 11 amino acid residues of the mature alpha-subunit, into the pMal-c2 vector of the MBP fusion expression system, and 3) expressing the fusion protein (MBP-cINA521) from the Escherichia coli and purifying it using affinity chromatography. In each trial, quail were randomly and equally assigned to one of two injection treatments as follows: 1) MBP-cINA521 in Freund's adjuvant, or 2) Freund's adjuvant (vehicular controls; CON). All immunizations were given subcutaneously and Freund's complete and incomplete adjuvant were used for primary and booster injections, respectively. In Trial 1, birds were given a primary challenge of 0.2 mg MBP-cINA521 per bird at 25 d of age, followed by booster immunizations (0.1 mg MBP-cINA521 per bird) at 33, 40, 47, 54 and 61 d of age and every 35 d thereafter. The CON birds received vehicular immunizations at the same time intervals. In Trial 2, birds treated with MBP-cINA521 received a primary challenge of 0.2 mg MBP-cINA521 per bird at 26 d of age, followed by booster immunizations (0.1 mg MBP-cINA521 per bird) using the same schedule as that used in Trial 1, with the exception that no boosters were given after 61 d of age. The CON birds received vehicular immunizations at the same time intervals. Collection of production performance data was initiated coincident with the laying of the first egg in each trial (i.e., beginning at 41 and 44 d of age for Trials 1 and 2, respectively) and continued for 30 1-wk periods of lay. Combined data from Trials 1 and 2 indicated that the mean +/- SE age at first egg lay was markedly decreased (P < 0.005) in MBP-cINA521-treated quail (53.4 +/- 0.9 d of age) when compared to the CON (57.6 +/- 1.3 d of age). Likewise, the mean +/- SE age at 50% egg production was reduced (P < 0.03) in quail immunized against inhibin (65.4 +/- 2.1 d of age) when compared to the CON (77.6 +/- 4.7 d of age). Total hen-day egg production was also higher (P < 0.05, Trial 1; P < 0.01, Trial 2) in MBP-cINA521-treated quail (88.7 +/- 1.4%, Trial 1; 90.1 +/- 1.2%, Trial 2) than in the CON birds (81.9 +/- 2.9%, Trial 1; 73.6 +/- 6.5%, Trial 2). Collectively, these findings provide evidence that inhibin immunoneutralization accelerated puberty and enhanced hen-day egg production during a 30-wk period of egg lay in Japanese quail.

摘要

在两项独立试验中,以麦芽糖结合蛋白-鸡抑制素α亚基521(MBP-cINA521)融合蛋白作为免疫原,评估了主动免疫抑制素对雌性日本鹌鹑(Coturnix coturnix japonica)生产性能的影响。融合蛋白MBP-cINA521由细菌麦芽糖结合蛋白(MBP)和鸡抑制素成熟α亚基的截短形式(cINA521)组成。MBP-cINA1521的构建方法如下:1)从鸡抑制素α亚基cDNA(cINA6;由P.A.约翰逊提供)中切下一个521 bp的PstI片段;2)将该片段(编码成熟α亚基除前11个氨基酸残基外的所有部分)克隆到MBP融合表达系统的pMal-c2载体中;3)从大肠杆菌中表达融合蛋白(MBP-cINA521),并使用亲和层析法进行纯化。在每项试验中,鹌鹑被随机且等分为以下两种注射处理之一:1)弗氏佐剂中的MBP-cINA521,或2)弗氏佐剂(载体对照;CON)。所有免疫均通过皮下注射进行,分别使用弗氏完全佐剂和不完全佐剂进行初次注射和加强注射。在试验1中,25日龄时每只鸡接受0.2 mg MBP-cINA521的初次免疫,随后在33、40、47、54和61日龄以及此后每35天进行加强免疫(每只鸡0.1 mg MBP-cINA521)。CON组鸡在相同时间间隔接受载体免疫。在试验2中,用MBP-cINA521处理的鸡在26日龄时每只接受0.2 mg MBP-cINA521的初次免疫,随后按照试验1的相同方案进行加强免疫(每只鸡0.1 mg MBP-cINA521),不同之处在于61日龄后不再进行加强免疫。CON组鸡在相同时间间隔接受载体免疫。在每项试验中,生产性能数据的收集在每只鸡产下第一枚蛋时开始(即试验1和试验2分别在41和44日龄开始),并持续30个为期1周的产蛋期。试验1和试验2的合并数据表明,与CON组(57.6±1.3日龄)相比,用MBP-cINA521处理的鹌鹑(53.4±0.9日龄)首次产蛋的平均±标准误年龄显著降低(P<0.005)。同样,与CON组(77.6±4.7日龄)相比,免疫抑制素的鹌鹑(65.4±2.1日龄)达到50%产蛋率的平均±标准误年龄降低(P<0.03)。用MBP-cINA521处理的鹌鹑的总母鸡日产蛋量也高于CON组鸡(试验1,P<0.05;试验2,P<0.01)(试验1为88.7±1.4%,试验2为90.1±1.2%)(试验1为81.9±2.9%,试验2为73.6±6.5%)。总体而言,这些发现提供了证据,表明抑制素免疫中和在日本鹌鹑30周的产蛋期内加速了青春期并提高了母鸡日产蛋量。

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