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Constructing and Screening Beta-Glucanase Activity of Metagenomic cDNA Expression Library of Digestive Gland of Achatina fulica
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| Author:
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MARIS KURNIAWATI, NUR HALIMAH, MUHAMMAD NUR HUDHA, SUDIYONO, PURKAN, SRI SUMARSIH, AFAF BAKTIR
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| Abstract:
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Digestive juice of Achatinafulica shows hydrolase activities which are useful as an antibiofilm. Insidethe digestive system
of snail many microorganisms live together as a normal flora.In this study, a metagenomicbased-activity technique was
carried out to construct the metagenomic expression library of digestive gland ofAchatinafulica. The metagenomic
library was screened for the recombinant harbouring genes encoding beta-glucanase again the specific substrate of
laminarin.The metagenomic cloning was carried out by using ?TriplEx2 as vector and E.coli XL1-Blue as host. The
longer than 0.5 cDNAs ligated to vector and the total ligation solution were 1000 µL.Circularization of each
recombinant phage inserted the target sequence were conducted in E.coli BM25.8, to become recombinant phagemids.
Recombinant phagemids screening were carried out by using restriction method to analyse the DNA insert.We
produced 2.8x108 plaques and 1.1x1010 library amplification titer. Seventeen hallo plaques from 100 µL of the ligation
solution positive screening with laminarin substrate.Eight recombinant phagemids were randomly picked and analysed
using restriction method showing different fragment. A high quantity cDNA library from digestive gland Achatinafulica
was successfully constructed.
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| Keyword:
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digestive system,recombinant harbouring genes,amplification titer
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2019.11.01.009
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| Download:
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