Gene Cloning Using Degenerate Primers and Genome Walking

Author(s):  
Javier A. Delgado ◽  
Steven Meinhardt ◽  
Samuel G. Markell ◽  
Rubella S. Goswami
2010 ◽  
Vol 36 (3) ◽  
pp. 526-532
Author(s):  
Ming-Xia WANG ◽  
Xiang GAO ◽  
Qi-Jiao CHEN ◽  
Jian DONG ◽  
Wan-Chun ZHAO ◽  
...  

2011 ◽  
Vol 34 (8) ◽  
pp. 1156-1164
Author(s):  
Yun-xia QIAN ◽  
Sun-xiao YANG ◽  
Hong LIANG ◽  
Lun QIAN ◽  
Kai-xian QIAN

BJHS Themes ◽  
2020 ◽  
Vol 5 ◽  
pp. 225-243
Author(s):  
Angela N.H. Creager

AbstractLaboratory instructions and recipes are sometimes edited into books with a wide circulation. Even in the late twentieth century, publications of this nature remained influential. For example, protocols from a 1980 summer course on gene cloning at Cold Spring Harbor Laboratory provided the basis for a bestselling laboratory manual by Tom Maniatis, Ed Fritsch and Joe Sambrook. Not only did the Molecular Cloning: A Laboratory Manual become a standard reference for molecular biologists (commonly called the ‘bible’), but also its recipes and clear instructions made gene cloning and recombinant DNA technologies accessible to non-specialists. Consequently, this laboratory manual contributed to the rapid spread of genetic-engineering techniques throughout the life sciences, as well as in industry. As is often the case with how-to books, however, finding a way to update methods in this rapidly changing field posed a challenge, and various molecular-biology reference books had different ways of dealing with knowledge obsolescence. This paper explores the origins of this manual, its publication history, its reception and its rivals – as well as the more recent migration of such laboratory manuals to the Internet.


2020 ◽  
pp. 1-9
Author(s):  
Nidia H. Montechiarini ◽  
Luciana Delgado ◽  
Eligio N. Morandi ◽  
Néstor J. Carrillo ◽  
Carlos O. Gosparini

Abstract During soybean seed germination, the expansive growth potential of the embryonic axes is driven by water uptake while cell wall loosening occurs in cells from the elongation zone (EZ). Expansins are regarded as primary promoters of cell wall remodelling in all plant expansion processes, with the expression profiles of the soybean expansins supporting their cell or tissue specificity. Therefore, we used embryonic axes isolated from whole seed and focused on the EZ to study seed germination. Using a suite of degenerate primers, we amplified an abundantly expressed expansin gene at the EZ during soybean embryonic axis germination, which was identified as EXP1 by in silico analyses. Expression studies showed that EXP1 was induced under germination conditions in distilled water and down-regulated by abscisic acid (ABA), which inhibits soybean germination by physiologically restraining expansion. Moreover, we also identified a time window of ABA responsiveness within the first 6 h of incubation in water, after which ABA lost control of both EXP1 expression and embryonic axis germination, thus confirming the early role of EXP1 in the EZ during this process. By contrast, EXP1 levels in the EZ increased even when germination was impaired by osmotically limiting the water availability required to develop the embryonic axes’ growth potential. We propose that these higher EXP1 levels are involved in the fast germination of soybean embryonic axes as soon as water availability is re-established. Taken together, our results show strong EXP1 expression in the EZ and postulate EXP1 as a target candidate for soybean seed germination control.


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