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Journal of Cell and Animal Biology Vol. 1(2), pp.024-028, September 2007
© 2007 Academic Journals
Full Length Research Paper
Isolation and cloning of
IGF-1R gene in bovine
Khairy MA Zoheir*1 and Li-Guo Yang1**
1College of Animal Science and Technology-
Huazhong Agricultural University- Wuhan 430070- Hubei Province- P R
China.
* National Research Center- Cell Biology Department- Dokki- Cairo-
Egypt.
**Corresponding author. E-mail:
yangliguo2006@yahoo.com.cn
Accepted
22 August, 2007
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Gene cloning, or
molecular cloning, has several different meanings to a molecular
biologist. A clone is an exact copy, or replica, of something. Molecular
biologists exploit the replicative ability of cultured cells to clone
genes. Recent advances
in the Restriction enzymes technologies have opened up new
opportunities in biotechnology and facilitate the way to isolate any
gene from any source of cells. Many types of restriction enzymes cleave
DNA away from their recognition site. By using the type III
restriction enzyme, we provide evidence to show that an intact
recognition site on the cleaved DNA sequesters the
restriction. We isolated and cloned the IGF-1 R gene from DNA by
subjecting DNA with one pair of restriction enzyme (EcoRI +
HindIII) and followed by ligation and transformation with the
suitable vector. The pair of restriction enzymes (EcoRI + HindIII)
which subjected to DNA template before the ligation process was the only
pair which had success to liberate IGF-1R fragment from the transformed
vector rather than other two pairs of restriction enzymes (EcoRI +
)
or (HindIII + )
although this two pair of restriction enzymes have recognition sites in
the vector. Moreover we have studied the effect of DNA template size on
the action of the restriction enzymes. Where less DNA size, more
efficient of the restriction enzyme action. This study is rapid and
useful new technology for isolation, cloning and sequencing the gene of
interest to be ready for further investigations as gene transfection
and/ or transfer.
Key words:
Restriction enzymes-
DNA technology- IGF-1R- bovine. |
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