文献类型： 外文期刊

作者： Liu, Xixi ¹ ; Xia, Dandan ² ; Luo, Jinjin ¹ ; Li, Mengyuan ² ; Chen, Lijuan ¹ ; Chen, Yiting ¹ ; Huang, Jie ¹ ; Li, Yanan ¹ ; Xu, Huayu ¹ ; Yuan, Yang ¹ ; Cheng, Yu ¹ ; Li, Zhiyong ¹ ; Li, Guanghao ¹ ; Wang, Shiyi ¹ ; Liu, Xinyong ¹ ; Liu, Wanning ¹ ; Zhang, Fengyong ¹ ; Liu, Zhichao ¹ ; Tong, Xiaohong ¹ ; Hou, Yuxuan ¹ ; Wang, Yifeng ¹ ; Ying, Jiezheng ¹ ; Ugli, Abdullaev Mirtemir Baxodir ³ ; Ergashev, Mukhammadjon Arabboevich ³ ; Zhang, Sanqiang ⁴ ; Yuan, Wenya ⁵ ; Xue, Dawei ⁶ ; Zhang, Jianwei ² ; Zhang, Jian ¹ ;

作者机构： 1.China Natl Rice Res Inst, State Key Lab Rice Biol & Breeding, Hangzhou 311400, Peoples R China

2.Huazhong Agr Univ, Natl Key Lab Crop Genet Improvement, Hubei Hongshan Lab, Wuhan 430070, Peoples R China

3.Rice Res Inst Uzbekistan, TASHKENT 111506, Uzbekistan

4.Hubei Univ Technol, Hubei Agr Machinery Engn Res & Design Inst, Wuhan 430068, Peoples R China

5.Hubei Univ, Sch Life Sci, State Key Lab Biocatalysis & Enzyme Engn, Wuhan 430062, Peoples R China

6.Hangzhou Normal Univ, Coll Life & Environm Sci, Hangzhou 311121, Peoples R China

关键词： barcode-indexed PCR; HiFi sequencing; protein-protein interactome; rice

期刊名称：ADVANCED SCIENCE （ 影响因子：14.1； 五年影响因子：15.6 ）

ISSN：

年卷期： 2025 年 12 卷 11 期

页码：

收录情况： SCI

摘要： Establishing the protein-protein interaction network sheds light on functional genomics studies by providing insights from known counterparts. However, the rice interactome has barely been studied due to the lack of massive, reliable, and cost-effective methodologies. Here, the development of a barcode-indexed PCR coupled with HiFi long-read sequencing pipeline (BIP-seq) is reported for high throughput Protein Protein Interaction (PPI)identification. BIP-seq is essentially built on the integration of library versus library Y2H mating strategy to facilitate the efficient acquisition of random PPI colonies, semi-mechanized dual barcode-indexed yeast colony PCR for the large-scale indexed amplification of bait and prey cDNAs, and massive pac-bio sequencing of PCR amplicon pools. It is demonstrated that BIP-seq could map over 15 000 high-confidence (approximate to 62.5% could be verified by Bimolecular fluorescence Complementation (BiFC)) rice PPIs within 2 months, outperforming the other reported methods. In addition, the obtained 23 032 rice PPIs, including 22,665 newly identified PPIs, greatly expanded the current rice PPI dataset, provided a comprehensive overview of the rice PPIs networks, and could be a valuable asset in facilitating functional genomics research in rice.