Gel dna purification kits

The full-length cDNA sequence of SaAACT and SaHMGS genes was isolated by 5′ and 3′ rapid amplification of cDNA ends (RACE)-PCR with the SMARTer RACE cDNA Amplification Kit (Clontech, Palo Alto, CA, USA) according to the manufacturer’s protocol. Gene-specific primers for 3′ RACE-PCR were designed on the basis of the initial data of AACT and HMGS unigenes in the S. album transcriptome^{71 (link)}. Based on data from the partial SaAACT and SaHMGS sequence generated from 3′ RACE-PCR, gene-specific primers were designed for 5′ RACE-PCR to obtain the remaining sequences of SaAACT and SaHMGS. PCR products were purified by gel DNA purification kits (Tiangen, Beijing, China) and ligated into the pMD18-T vector (Takara Bio Inc., Dalian, China). Recombined plasmids were transformed into DH5α-competent E. coli cells (Takara Bio Inc., Dalian, China) and sequenced.

Mueller Hinton (M-H) broth and de Man, Rogosa, and Sharpe (MRS) broth were purchased from Becton Dickenson (United States). Biochemical reagents (i.e. arabinose, pyruvate, tellurite, arginine, glucose, inulin, lactose, mannitol, maltose, melibiose, raffinose, ribose, sucrose, sorbitol, sorbose, trehalose, and xylose) and antimicrobial agents (i.e. ampicillin, chloramphenicol, ciprofloxacin, clarithromycin, erythromycin, gentamicin, nitrofurantoin, norfloxacin, tetracycline and vancomycin) were obtained from Hangzhou Microbe Reagent Co., Ltd. (Hangzhou, China). Primers were synthesized by the Tsingke Biotechnology Limited Company (Wuhan, China). Taq DNA Polymerase and deoxynucleotide triphosphates (dNTPs) were purchased from Takara (China). Defibrinated rabbit blood was purchased from Zhengzhou Kowloon Biological Products Co., Ltd. (Zhengzhou, China). DNA isolation kits and gel DNA purification kits were purchased from Tiangen Biotech (Beijing, China).