The whole genome sequences of G. lemaneiformis were captured from our own laboratory database [38 (link)]. For catching the PGM protein sequence, the Hidden Markov Model (HMM) analysis was used for the search. The HMM profiles of the phosphoglucomutase/phosphomannomutase (PF02878, PF02879, PF02880 and PF00408) from the Pfam protein family database Pfam version 35.0 (http://pfam.xfam.org/ , 29 June 2022) were used as the query to search the G. lemaneiformis protein database with an e-value ≤ e−10. Blastp was subsequently used to search for missing possible PGM candidates. Pfam and SMART (http://smart.embl-heidelberg.de/ , 29 June 2022) were then used to confirm the conserved domain, and the protein sequences lacking the conserved domains were excluded.
The total RNA from G. lemaneiformis was isolated by a RNeasy Plant mini kit (QIAGEN, Germany), according to the manufacturer’s protocol. The cDNA for the full-length sequence cloning was subsequently synthetized by using HiScript II Reverse Transcriptase Kit (Vazyme, Nanjing, China). The opening reading frames (ORFs) of three possible GlPGM were annotated as GlPGM1, GlPGM2 and GlPGM3, and finally amplified by PCR, using Prime Star Kit (Takara, Dalian, China). The gene-specific primers with different restriction endonuclease site are indicated in the additional fileTable S1 (Supplementary Materials) .
The total RNA from G. lemaneiformis was isolated by a RNeasy Plant mini kit (QIAGEN, Germany), according to the manufacturer’s protocol. The cDNA for the full-length sequence cloning was subsequently synthetized by using HiScript II Reverse Transcriptase Kit (Vazyme, Nanjing, China). The opening reading frames (ORFs) of three possible GlPGM were annotated as GlPGM1, GlPGM2 and GlPGM3, and finally amplified by PCR, using Prime Star Kit (Takara, Dalian, China). The gene-specific primers with different restriction endonuclease site are indicated in the additional file
Free full text: Click here
