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本帖最后由 细胞海洋 于 2011-10-11 09:11 编辑
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! W: U/ ]9 M) O3 I+ gThe discovery of DNA as the genetic material brought great hope to scientists all over the, U1 |' K: q |1 y) d
world. It was believed that many of the lingering questions in genetics and the mechanisms
; B. h) m* v/ N( K4 W9 Fof heredity would finally be answered. However, as often is the case in science, more questions# V$ d0 z0 @! R$ ~
arose out of this discovery. What defines a gene? What are the mechanisms of gene
& x; D; t7 v; t9 k9 s' w3 G/ |regulation? Further discovery and technological innovations brought about sequencing3 N, m; J: F; J' k5 g4 m* q, P
techniques that allowed the study of complete genomes from many organisms, including
& Y7 g( q+ b, s& t8 [9 NArabidopsis and humans. Despite all the excitement surrounding these technologies, many
7 G+ H" b3 ` D( a' Lfeatures of the genome remained unclear. Peculiar characteristics in genome composition
* a' p& z8 }, ]" O. {0 r7 vsuch as significant redundancy consisting of many repetitive elements and noncoding& b; W) x% Y; K! h/ J3 @
sequences, active transcriptional units with no protein product, and unusual sequences in1 J" [, S/ b8 x
promoter regions added to the mysteries of genetic make-up and gene regulation. Indeed,
C- x, K; }0 \( @$ Vthe more we discovered about the genome, the more difficult it became to understand the
! N& ]0 V7 K! u5 f: Ecomplexity of cellular function and regulation.
o- [6 ^' Q1 ^' |Out of the study of the intricacies of the genome and gene regulation, arose a new; V" ^$ N5 K3 p( ?3 I8 B! Q7 o7 n
science that was independent of actual DNA changes, but critical in maintaining gene
1 s( W: { {; j) @* Q2 F- Rregulation and genetic stability. Epigenetics, literally translated as “above genetics,” is the
3 P$ f O/ t J$ Fscience that describes the mechanisms of heritable changes in gene regulation that does
5 C- X! a# F, L2 X2 F/ Q* enot involve modifications of DNA sequence. These changes may last through somatic cell2 s* I; Y5 W$ C$ l
division and, in some cases, throughout multiple generations.
* d9 ]2 F7 g' J0 J$ J1 xEpigenetics is perhaps one of the most popular and quickly evolving fields of modern
0 T2 q( z: Z/ Y A' o8 m. M+ a9 @$ Ascience. Despite the fact that the ideas behind epigenetics had already been developing in
3 ~1 ?8 M& Y7 [' f& ?the late nineteenth and early twentieth centuries, major advances have only occurred
/ Z. F: A N7 m6 t- ?" w; Fwithin the last 10–15 years as the mechanisms surrounding epigenetic regulation began to \; s! |* D% @6 P6 a* ?% {
be uncovered. It was hoped by many that the mysteries of gene regulation and inheritance9 |+ r; x% B- v( L
that remained unanswered would finally be elucidated with the help of this new science.
6 A( p* j. _, n2 H3 L2 \7 RSince, the understanding of the contribution of epigenetic regulation to cell function has$ b) w7 D# m9 N$ Y
helped scientists from many distinct fields of research such as molecular biology, population
8 g5 l6 u/ ?6 E n Egenetics, microbiology, ecology, developmental biology, and evolution.
( ~* f- b; m( Q- wGene silencing as an epigenetic mechanism to control gene expression was first% |# m* d4 l$ ` P0 E
described in plants. This occurred with the beginning of the era of plant transgenesis, and
4 c1 g) f. Z: G9 u. q6 r; _almost undermined the new paradigm of improvement of plant performance via transgenic+ R4 b# ]% r0 d
techniques. Silencing was a serendipitous discovery, as this finding revitalized the3 ~5 m4 q) U) Z/ i" C0 o6 b& o
field of epigenetics. Phenomena such as plant acclimation and adaptation to stress, hybrid L" o" B, }9 j% {1 L1 ?$ r9 ~& H0 y
and heterozygote vigor (heterosis), plant tolerance to viral infection, transgenerational% n5 z3 }5 `3 U. p1 R
changes in genome stability, paramutations, among others, are now considered excellent. \9 t" w9 j- s! H2 I
candidates for regulation via epigenetic mechanisms. Future studies involving various protocols
& z' B q" g9 ]0 R- ofor the analysis of methylation patterns, histone modifications, chromatin structure,9 c ?% D4 [2 O. A% N4 q
and small RNA expression, the hallmarks of epigenetic regulation, will undoubtedly help- i8 i1 c7 V4 L c# N8 m- y
to explain these phenomena. It will be exciting to discover how plants utilize these mechanisms& e2 e5 X, ?7 ~1 Y4 ?. e/ L
to adapt to stress, and how we can manipulate these characters for the generation of
/ Y7 K; @9 g0 ?; q+ U/ {1 l8 lbetter and hardier crops.' o- U K; N5 n* t7 T7 @; n# ?
vi Preface
/ w; d `; i! w! y; V) qIn this book we have collected a variety of protocols for the study of the function of
( u4 I& o0 b+ T/ Zsmall noncoding RNAs, DNA methylation, and histone modifications in plants. Where possible: H" @9 `* W h8 C0 M& r4 n" k
and appropriate, we presented several protocols with different degrees of complexity.: a. g8 ~+ y2 a, [
We also include protocols for plant transgenesis and the analysis of genome stability, with a: m, v8 e7 l% T* K+ _8 [
discussion for their applications to epigenetic studies. It was our aim to put together a single- O5 j; F; K% o* s4 v+ L& G
manual that researchers in the field of plant epigenetics can turn to in hopes to answer the
5 [; m; ^$ q( i) qmany yet undiscovered and unexplained phenomena in plant biology.
m- C- ]" P- ?3 n: P8 ULethbridge, AB, Canada Igor Kovalchuk2 L$ v- h- e; Y q& t: H
Franz J. Zemp
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