PCR Amplification and Molecular Cloning of DNA Encoding Betaine-aldehyde Dehydrogenase; Its ., Role in Salinity and Drought Tolerance From the i I'I I Halophyte Arthrocnemon glaucum
Amr Shahan Abdelghany;
Abstract
Environmental stresses affect plant growth, development, and hence crop productivity. Salt stress is a member of water stress factors. A secondary stress is following salt stress, osmotic stress. Ostnotic stress makes cell loose turgor and in turn causing plasmolysis. Osmoregulation is the defense mechanism against osmotic injury, which is the maintenance of cell turgor by a sufficient increase in cell so'! utes to compensate for the external osmotic stress. This can be achieved by certain molecules that work as compatible solutes (osmolyte).
As a powerful osmoprotective osmolyte, betaine is accumulaied in the cytoplasm of many plants as well as in a vruiety of organisms. Betaine-aldehyde dehydrogenase (BADH EC 1.2.1.8) catalyzes the last step in the betaine biosynthetic pathway. BADH gene has been cloned and sequenced, one from spinach and two from sugru• beet.
Halophytes are capable to survive best at extremely salt-stress
conditions. Arthrocnemon glaucum, a chenopod inhabits the North W'
coast of Egypt. It can survive salinity stress up to 800 mM NaCI.
est
...
In our work, polymerase chain reaction (PCR) technique was used in amplification of specific DNA region. As a step towards cloning of its full length, DNA encoding BADH was amplified by polymerase c'hain reaction (PCR) from the halophyte Arthrocnemon glaucum DNA.
Amplification was accomplished by using two sets of primers. The first was degenerate outer set deduced from a highly conserved boll that was revealed from amino acid sequence alignment of BADH with' nine different dehydrogenases. The second nested set was specific for BADH deduced from the alignment of three BADH genes. The nested amplified product was cloned in pBluescript vector. The authenticity of the clone was confmned by DNA hybridization witl1 a specific probe that was cloned from spinach. The spinach probe was cloned by reverse transcriptase (RT-PCR) using two successive PCR primers sets specific
for BADH. One set amplified a region of 1.326 kilobases of DNA
As a powerful osmoprotective osmolyte, betaine is accumulaied in the cytoplasm of many plants as well as in a vruiety of organisms. Betaine-aldehyde dehydrogenase (BADH EC 1.2.1.8) catalyzes the last step in the betaine biosynthetic pathway. BADH gene has been cloned and sequenced, one from spinach and two from sugru• beet.
Halophytes are capable to survive best at extremely salt-stress
conditions. Arthrocnemon glaucum, a chenopod inhabits the North W'
coast of Egypt. It can survive salinity stress up to 800 mM NaCI.
est
...
In our work, polymerase chain reaction (PCR) technique was used in amplification of specific DNA region. As a step towards cloning of its full length, DNA encoding BADH was amplified by polymerase c'hain reaction (PCR) from the halophyte Arthrocnemon glaucum DNA.
Amplification was accomplished by using two sets of primers. The first was degenerate outer set deduced from a highly conserved boll that was revealed from amino acid sequence alignment of BADH with' nine different dehydrogenases. The second nested set was specific for BADH deduced from the alignment of three BADH genes. The nested amplified product was cloned in pBluescript vector. The authenticity of the clone was confmned by DNA hybridization witl1 a specific probe that was cloned from spinach. The spinach probe was cloned by reverse transcriptase (RT-PCR) using two successive PCR primers sets specific
for BADH. One set amplified a region of 1.326 kilobases of DNA
Other data
| Title | PCR Amplification and Molecular Cloning of DNA Encoding Betaine-aldehyde Dehydrogenase; Its ., Role in Salinity and Drought Tolerance From the i I'I I Halophyte Arthrocnemon glaucum | Other Titles | التضاعف باستخدام التفاعل السلسى للبوليميراز والاستنسال الجزيئى للمحض النووى المسئول عن انزيم بيتان الدهيد دهيدروجيناز ودوره فى تحمل المللوحة والجفاف والمعزول من نبات ارثوكنيمون جلوكم المحب للملوحة | Authors | Amr Shahan Abdelghany | Issue Date | 1995 |
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