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|Title: ||Molecular cloning of hypoxia induced genes barnacle larvae|
|Other Titles: ||Di yang you dao zhi teng hu you ti ji yin de fen zi kelong|
|Authors: ||So, Chun Hung (蘇振雄)|
|Department: ||Department of Biology and Chemistry|
|Degree: ||Master of Philosophy|
|Issue Date: ||2004|
|Publisher: ||City University of Hong Kong|
|Subjects: ||Barnacles -- Effect of Anoxemia on|
|Notes: ||CityU Call Number: QL444.C58 S6 2004|
Thesis (M.Phil.)--City University of Hong Kong, 2004
Includes bibliographical references (leaves 124-135)
ix, 136 leaves : ill. (some col.) ; 30 cm.
|Abstract: ||Hypoxia is an environmental stress that will trigger a series of gene expression for an
organism to overcome it. Barnacle (Balanus amphitrite), as a cosmopolitan species,
plays an important role in the ecosystem. In the current study, molecular response to
hypoxia is investigated in barnacle larvae. Barnacle larvae in the nauplii stage were
treated in hypoxic condition (1 mg Oz/L) for 24 hours and the induction/repression of
gene expressions were studied by RAP (RNA fingerprinting by arbitrary primed PCR).
Thirty-nine differential PCR products were obtained from 46 arbitrary chosen primers.
After cloning and sequencing, 34 products showed no homolog in the GenBank
database and, therefore, they were novel genes of barnacle. The remaining 5 were
identified to heat shock protein 70 (hsp-70), cell division factor, p76 protein and a
protein kinase. To get the full length sequence of these genes, RACE (Rapid
amplification of cDNA ends) was used, with gene specified primers designed from the
RAP amplified products. To amplify unique single products, optimization of the
concentration of template and annealing temperature of primer was performed for
each set of primers. The RACE amplified products were subcloned and sequenced.
Except protein kinase, overlapping sequence of 5’ RACE and 3’ RACE products were
obtained The length of hsp-70, cell division factor and p76 gene were 2320 bp, 3055
bp and 1744 bp respectively. Deduced amino acid sequences showed high
similarity to human homolog. Northern hybridization was used to investigate the
gene expression of these genes in hypoxic adult barnacle. The results showed that
hypoxic treatment of 1.0 mg O2/L for 24 hours in barnacle adults induced a 2.5 fold
increase in HSP70 transcript. Whole-mount in situ hybridization of hsp70 in nauplii
larvae showed the increase in the expression of hsp70 gene after hypoxia treatment.
These results demonstrated that hsp70 gene expression was induced by hypoxia in the
both adult and larva of barnacle, revealing a common response in gene expression to
|Online Catalog Link: ||http://lib.cityu.edu.hk/record=b1871674|
|Appears in Collections:||BCH - Master of Philosophy |
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