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Increasing transient expression of CAT gene in Porphyra haitanensis by Matrix attachment regions and 18S rDNA targeted homologous recombination
Author(s) -
Zuo Zhenghong,
Li Bowen,
Wang Chonggang,
Cai Jiali,
Chen Yixin
Publication year - 2007
Publication title -
aquaculture research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 89
eISSN - 1365-2109
pISSN - 1355-557X
DOI - 10.1111/j.1365-2109.2006.01645.x
Subject(s) - biology , microbiology and biotechnology , 18s ribosomal rna , transformation (genetics) , gene , homologous recombination , ribosomal rna , genetics
To test whether matrix attachment regions (MARs) and 18S rDNA can influence CAT gene transient expression positively in the red algae Porphyra haitanensis , a targeting vector pHR‐CAT containing a portion of the 18S rDNA from P. haitanensis , pMAR1‐HR‐CAT containing one MAR from silkworm and a portion of the 18S rDNA from P. haitanensis and pMAR2‐HR‐CAT containing two MARs from silkworm and a portion of the 18S rDNA from P. haitanensis were constructed. With the electroporation method, the vectors were transferred into the protoplasts from the thalli of P. haitanensis . The results showed that the expression of chloramphenicol acetyl transferase (CAT) protein in transformed cells reached a maximum at 96 h after transformation. It was increased markedly with the pMAR2‐HR‐CAT compared with the pHR‐CAT or the pCAT @ 3‐control vector ( P <0.01), and it was increased inconspicuously with pHR‐CAT compared with the pCAT @ 3‐control vector ( P >0.05). It is suggested that MAR from silkworm could enhance the transient expression of foreign genes in P. haitanensis .