Abstract: In order to construct a recombinant attenuated Salmonella typhimurium type Ⅲ secretion system which carries the hemagglutinin‑neuraminidase（HN） protein gene of Newcastle disease virus（NDV），the HN gene was amplified from the plasmind pMD18‑T‑HN and cloned into the vector pYA3493‑sopE_Nt100.Then the recombinant vector pYA3493‑sopE_Nt100‑HN was transformed into attenuated Salmonella typhimurium strain Δ crpΔ asd SL1344 by electroporation，and the growth characteristics，genetic stability，expression characteristics and virulence of the recombinant strain Δ crpΔ asd SL1344（pYA3493‑sopE_Nt100‑HN）were analyzed. The identification results of PCR and enzyme digestion showed that the recombinant attenuated Salmonella typhimurium strain Δ crp Δ asd SL1344（pYA3493‑sopE_Nt100‑HN）was successfully constructed. The growth curve of the recombinant strain was similar to that of the complementary strainΔ crpΔ asd SL1344（pYA3493‑sopE_Nt100）and the deletion strain Δ crpΔ asd SL1344，which were significantly different from their parent strain SL1344.The HN gene was stably inherited in the recombinant strain.The HN protein was presented by the attenuated Salmonella typhimurium Ⅲ type secretion system and expressed in chicken embryo fibroblast（CEF）.The LD50 of the recombinant strain was not significantly different from the complementary strain and the deletion strain，but significantly different from the parent strain SL1344. The above results indicated that the recombinant attenuated Salmonella typhimurium Ⅲ secretion system carrying the NDV HN gene was successfully constructed，which had high safety，genetic stability，and expression efficacy of the NDV HN gene.

Key words: Newcastle disease virus, HN gene, Type Ⅲ secretory system, Attenuated Salmonella, Biological characteristics

摘要： 为构建能稳定携带新城疫病毒（Newcastle disease virus，NDV）血凝素-神经氨酸酶蛋白（Hemagglutinin‑neuraminidase protein，HN）基因的减毒鼠伤寒沙门菌Ⅲ型分泌系统，从pMD18-T-HN 中扩增出NDV HN 基因，并将其克隆入载体pYA3493-sopE_Nt100 ，构建重组载体pYA3493-sopE_Nt100-HN，然后将其电转化至减毒鼠伤寒沙门菌Δ crpΔ asd SL1344，并对其重组菌的生长特性、遗传稳定性、表达特性和毒力特性等进行分析。结果显示，携带NDV HN 基因的重组减毒鼠伤寒沙门菌Δ crpΔ asd SL1344（pYA3493-sopENt100-HN）构建成功；重组菌Δ crpΔ asd SL1344（pYA3493-sopE_Nt100-HN）的生长趋势与互补菌ΔcrpΔasd SL1344（pYA3493-sopE_Nt100）和缺失菌Δ crpΔ asd SL1344相似，与亲本菌株SL1344有明显差异；重组菌能够稳定表达HN 基因；减毒鼠伤寒沙门菌Ⅲ型分泌系统能够有效递呈HN 基因，并诱导其在鸡胚成纤维细胞（Chicken embryo fibroblast，CEF）内表达；重组菌的LD50与互补菌和缺失菌没有明显差异，与亲本菌株SL1344差异较大。以上结果表明，成功构建了携带NDV HN 基因的新型减毒鼠伤寒沙门菌Ⅲ型分泌系统Δ crpΔ asd SL1344（pYA3493-sopENt100-HN），能够有效递呈NDV HN 基因并使其高效表达，且安全性高，遗传稳定。

关键词: 新城疫病毒, HN 基因, Ⅲ型分泌系统, 减毒沙门菌, 生物学特性