title: [Construction of suppression subtractive hybridization (SSH) library of copepod Pseudodiaptomous annandalei and its ferritin cDNA cloning and differential expression under nickel stress].
abstract: To study the molecular response mechanisms of copepod to nickel stress, a suppression subtractive hybridization (SSH) cDNA library of Pseudodiaptomous annandalei under nickel stress was constructed by using SSH technique, and a total of 140 clones were randomly picked from the growing colonies and identified by PCR. The recombinant rate of the library was 98.6%, and the volume of the library was 1.12 x 10(6) cfu. After the recombinant plasmids were sequenced, a partial cDNA fragment of ferritin was recognized based on BLAST searches in NCBI, with a size of 859 bp and continuously encoding 170 amino acid residues. The semi-quantitative PCR results showed that the ferritin cDNA under 24 h nickel stress was distinctly up-regulated. The successful construction of the SSH library and the obtaining of ferritin cDNA fragment would supply basis for the further study of the molecular response mechanisms of copepod to nickel stress.
authors.affiliation: State Key Laboratory of Marine Environmental Science, College of Ocean & Earth Sciences, Xiamen University, Xiamen 361005, Fujian, China. jielanjiang@gmail.com
authors.forename: Jie-Lan
authors.initials: JL
authors.lastname: Jiang
authors.forename: Gui-Zhong
authors.initials: GZ
authors.lastname: Wang
authors.forename: Li-Sheng
authors.initials: LS
authors.lastname: Wu
authors.forename: Shao-Jing
authors.initials: SJ
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D033342:Copepoda / Q000235:genetics* / Q000502:physiology*; D005293:Ferritins / Q000235:genetics*; D015723:Gene Library; D008969:Molecular Sequence Data; D009532:Nickel / Q000494:pharmacology*; D009693:Nucleic Acid Hybridization; D013312:Stress, Physiological; D059467:Transcriptome
chemical_list: D009532:Nickel; D005293:Ferritins