February 15-18, 2009 Washington State Convention Center Seattle ...

ISOLATION, IDENTIFICATION AND BIOLOGICAL FUNCTION OF A PUTATIVE SHRIMP
β-Actin Promoter from Litopenaus Vannamei
Yuanan Lu, Marcus Soderlund and Chengxiang Wu
Department of Public Health Sciences
University of Hawaii
Honolulu, Hawaii 96822 USA
ylu@pbrc.hawaii.edu
Shrimp farming has become an establishing industry in many countries over the world. However, viral diseases have proven
to be a serious threat to shrimp farming and pathogenic viruses are frequently reported to cause 100% shrimp mortality and
tremendous economic losses worldwide. There are currently no effective methods to treat viral diseases. The use of molecular
biology techniques to produce pathogen resistant strains of shrimp through genetic transformation technology is considered a
highly promising strategy for control of shrimp viral diseases. A major challenge to current development of such biotechnology
is the lack of a suitable host-derived promoter to drive the effective expression of desirable genes in shrimp. To meet this urgent
need for the development of transgenic shrimp, we have recently employed molecular cloning and sequencing techniques
and isolated and identified a putative promoter gene from litopenaeus shrimp (L. vannamei), shrimp β-actin promoter. This
represents the first promoter gene identified and reported from crustacean species. Biological function of newly discovered
shrimp promoter has been demonstrated using a newly constructed gene expression cassette system under the control of such
shrimp promoter. Experimental tests have confirmed high level of stable expression of red fluorescent protein (RFP), a reporter
transgene, and also an antiviral gene of shrimp white spot syndrome virus in a variety of in vitro cell cultures derived from both
mammalian and marine animal species using fluorescent microscopy, Western blot and enzyme-linked immunosorbent assay.
With the previously established jetPEI/DNA technique for gene delivery into the shrimp zygotes, this putative shrimp promoter
forms the essential base for genetic shrimp transformation technology and makes it possible for the development and production
of viral-resistant transgenic shrimp strains for world shrimp aquaculture.
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