Escherichia coli strain dh5α

In this study, the Brucella abortus 2308 (B. abortus) wild strain (WT) and 2308 ΔbspF mutant strain were obtained from the Institute of Military Sciences (Beijing, China) and were grown in tryptic soy agar (TSA; Takara, Kusatsu, Japan) and tryptic soy broth (TSB; Takara, Kusatsu, Japan).
The Escherichia coli strain DH5α (TransGen Biotech) was cultured in Luria–Bertani (LB) medium for gene cloning. HEK-293T cells, derived from human embryonic kidney cells, and HeLa cells, derived from cervical cancer cells, both from our laboratory, were cultured in Dulbecco’s minimal essential medium (DMEM) containing 10% fetal bovine serum (FBS) (Gemin, Woodland, CA, USA) at 37 °C and 5% CO₂.

The pRS300 vector, expression vector pBI121 (CaMV 35S promoter, resistance marker Kanamycin) and Agrobacterium tumefaciens GV3101 are preserved by Genetic Laboratory, College of Life Science and Technology, Gansu Agricultural University (Lanzhou, China). Rapid Plant Genomic DNA Isolation Kit, SanPrep Column Plasmid Mini-Preps Kit, DiaSpin DNA Gel Extraction Kit, One Step qRT-PCR Probe Kit and SuperReal PreMix Plus (SYBR Green) were purchased from Sangon Biotech (Shanghai, China) Co., Ltd. TRNzol Universal Reagent and MiRNA cDNA First Strand Synthesis Kit were purchased from Accurate Biology (Hunan, China) Co., Ltd. pMD18-T vector, restriction endonuclease, T4 DNA ligase, 2 × EasyTaq^® PCR SuperMix and Escherichia coli strain DH5α were purchased from TransGen Biotech (Beijing, China) Co., Ltd. Takara EX Taq(#RR902A), SMART ™ RACE cDNA Amplification Kit, Thiobarbituric acid (TBA), Nitro-Blue Tetrazolium (NBT), 2-methoxyphenol and other reagents were purchased from Takara Bio (Dalian, China) Co., Ltd. Primer synthesis and sequencing was completed by Lanzhou Tqgene Gene Biotechnology Co., Ltd.

T. asperellum CAU126 was screened, identified, and preserved in China General Microbiological Culture Collection Center (CGMCC No. 3.5921). Escherichia coli strain DH5α (TransGene, Beijing, China) was employed as the host for the cloning and sequencing of TaproA1. The K. phaffii GS115 (his4, Mut⁺, Invitrogen) strain was the chassis host for TaproA1 expression. pEASY-Blunt (TransGen, Beijing, China) and pPIC9K (Invitrogen, Carlsbad, CA, USA ) plasmids were utilized as the cloning and the expression vectors, respectively. FastPfu DNA polymerase, NEBbuilder® HiFi DNA Assembly Master Mix, and restriction enzymes (NEB, Frankfurt, Germany) were used for DNA manipulation. Duck blood hemoglobin and plasma protein were obtained from Handan Xinheng Biotechnology Co., Ltd. Casein sodium salt from bovine milk was purchased from Sigma-Aldrich (St. Louis, MO, USA), and all other reagents used herein were commercially accessible and analytical grade.