Cloning, characterization, and expression of Cytochrome b (Cytb)—a key mitochondrial gene from Prorocentrum donghaiense

Liyuan Zhao; Tiezhu Mi; Yu Zhen; Zhigang Yu

doi:10.1007/s00343-012-1137-4

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Cloning, characterization, and expression of Cytochrome b (Cytb)—a key mitochondrial gene from Prorocentrum donghaiense

Updated：2023-05-16

- Cloning, characterization, and expression of Cytochrome b (Cytb)—a key mitochondrial gene from Prorocentrum donghaiense
- Journal of Oceanology and Limnology Vol. 30, Issue 3, Pages: 424-432(2012)
- Affiliations：
  
  1. Key Laboratory of Marine Environment and Ecology Ministry of Education Ocean University of China
  2. Third Institute of Oceanography State Oceanic Administration
  3. Key Laboratory of Marine Chemistry Theory and Technology Ministry of Education Ocean University of China
- Author bio：
- Funds：
- DOI：10.1007/s00343-012-1137-4
  CLC：
- Online First：19 September 2011，
  
  Published：2012
- Accepted：
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Zhao, L., Mi, T., Zhen, Y. et al. Cloning, characterization, and expression of Cytochrome b (Cytb)—a key mitochondrial gene from Prorocentrum donghaiense, ., Chin. J. Ocean. Limnol. 30, 424–432 (2012). https://doi.org/10.1007/s00343-012-1137-4
DOI：

Zhao, L., Mi, T., Zhen, Y. et al. Cloning, characterization, and expression of Cytochrome b (Cytb)—a key mitochondrial gene from Prorocentrum donghaiense, ., Chin. J. Ocean. Limnol. 30, 424–432 (2012). https://doi.org/10.1007/s00343-012-1137-4 DOI：

摘要

Abstract

Mitochondrial cytochrome b (Cytb)

one of the few proteins encoded by the mitochondrial DNA

plays an important role in transferring electrons. As a mitochondrial gene

it has been widely used for phylogenetic analysis. Previously

a 949-bp fragment of the coding gene and mRNA editing were characterized from Prorocentrum donghaiense

which might prove useful for resolving P. donghaiense from closely related species. However

the full-length coding region has not been characterized. In this study

we used rapid amplification of cDNA ends (RACE) to obtain full-length

1 124 bp cDNA. Cytb transcript contained a standard initiation codon ATG

but did not have a recognizable stop codon. Homology comparison showed that the P. donghaiense Cytb had a high sequence identity to Cytb sequences from other dinoflagellate species. Phylogenetic analysis placed Cytb from P. donghaiense in the clade of dinoflagellates and it clustered together strongly with that from P. minimum. Based on the full-length sequence

we inferred 32 editing events at different positions

accounting for 2.93% of the Cytb gene. 34.4% (11) of the changes were A to G

25% (8) were T to C

and 25% (8) were C to U

with smaller proportions of G to C and G to A edits (9.4% (3) and 6.2% (2)

respectively). The expression level of the Cytb transcript was quantified by real-time PCR with a TaqMan probe at different times during the whole growth phase. The average Cytb transcript was present at 39.27±7.46 copies of cDNA per cell during the whole growth cycle

and the expression of Cytb was relatively stable over the different phases. These results deepen our understanding of the structure and characteristics of Cytb in P. donghaiense

and confirmed that Cytb in P. donghaiense is a candidate reference gene for studying the expression of other genes.

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