小菜蛾清道夫受体基因家族鉴定与生物信息学分析

【目的】小 菜蛾（Plutella xylostella）对化学杀虫性抗药性逐年上升，使 RNAi （RNA interference）农药广受关注。小菜蛾清道夫受体 （Scavenger receptor, SR）在细胞免疫中表现出吞噬和清除病原体的能力，使其成为 RNAi 农药的备选靶标基因。为确定小菜蛾 SR 基因相关靶标，鉴定分析了小菜蛾 SR 基因家族成员信息。【方法】基于全基因组数据，对小菜蛾 SR 基因家族的理化性质、跨膜结构、系统发育进化、蛋白质二三级结构预测、蛋白保守基序、磷酸化位点预测等内容进行分析。【结果】 小菜蛾 SR 基因家族有 12 个 CD36 蛋白序列，具有相似的理化性质和功能结构，均含有多个亲水性和疏水性区域及 2 个跨膜螺旋区，蛋白质二、三级结构的主要特征为无规则卷曲，磷酸化位点中丝氨酸含量最高。小菜蛾与家蚕（Bombyx mori）、冈比亚按蚊（Anopheles gambiae）、黑腹果蝇（Drosophila melanogaster）、烟草天蛾（Manduca sexta）、赤拟谷盗（Tribolium castaneum）等 6 个物种共包含 117 条 CD36 蛋白序列，这些序列在进化树上分成 5 个分支，具有相似结构的基因聚为同一支，进化保守性强。小菜蛾 CD36px4、CD36px5 具有更复杂的三级结构，且 motif19、motif20 两个基序只存在 于 CD36px4，推测其在物种进化中发挥某些特异功能，CD36px4 磷酸化位点数量多达 143 个，可能与凋亡细胞的吞噬和清除功能表达有关。【结论】鉴定分析到 12 个小菜蛾 SR 蛋白均为跨膜型蛋白，且含有丰富的磷酸化位点，预测其可参与细胞内外物质传输、信号传递及能量代谢等机体活动，具有介导细胞凋亡、吞噬和清除凋亡细胞的功能，可作为 RNAi 农药的备选靶标，为小菜蛾 RNAi 农药靶标的筛选提供重要依据。     

猪繁殖与呼吸综合征抗病育种研究进展

猪繁殖与呼吸综合征(porcine reproductive and respiratory syndrome, PRRS)是由猪繁殖与呼吸综合征病毒(PRRS virus, PRRSV)引起的一种高度接触性传染病，严重危害我国乃至世界养猪业。然而由于PRRSV抗原的多变性，目前包括疫苗接种、药物治疗等在内的防治措施效果不佳。因此，随着现代分子生物学技术的不断发展，基于基因编辑技术对猪PRRS的抗病育种逐渐发展起来。本文简述了PRRS的临床症状，重点回顾了国内外PRRS抗病育种研究进展，通过分析PRRS的致病机制，重点阐述了PRRSV受体及针对不同受体进行编辑的体内及体外抗病毒效果，以期为未来深入研究PRRSV致病机制、开发PRRS抗病品种提供理论依据。     

Pig macrophages with site-specific edited CD163 decrease the susceptibility to infection with porcine reproductive and respiratory syndrome virus

Porcine reproductive and respiratory syndrome (PRRS) is recognized as one of the most infectious viral diseases of swine. Although Cluster of differentiation 163 (CD163) is identified as an essential receptor for mediating PRRS virus (PRRSV) infection, the important residues involved in infection on CD163 are still unclear. Therefore, it is very important to identify these key residues to study the mechanism of PRRSV infection and to generate anti-PRRSV pigs. In this study, we first generated immortalized porcine alveolar macrophage (IPAM) cell lines harboring 40-residues (residues 523–562, including R561 (arginine (R) at position 561)) deletion of CD163. PRRSV infection experiments showed that these IPAM cell lines were completely resistant to PRRSV infection. We then generated cloned pigs carrying CD163-R561A (an arginine (R) to alanine (A) substitution at position 561 of CD163). PRRSV challenge experiments in porcine alveolar macrophages (PAMs) isolated from the CD163-R561A pigs showed significantly lower susceptibility to PRRSV than that of CD163-R561 PAMs. Through this study, we show that CD163 523–562 contains essential residues for mediating PRRSV infection, and that CD163 R561 significantly contributes to PRRSV infection but is not essential for infection. These functional sites can therefore serve as new targets for understanding the mechanism of PRRSV infection. Furthermore, CD163-R561A pigs can be used as an important model for improving pig germplasm with resistance against PRRSV.