2004 Fiscal Year Final Research Report Summary
Analysis and detecction of polymorphism of palindrome complex on Y chmmosome in idiopathic male infertility
| Project/Area Number |
15591677
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Urology
|
|---|
| Research Institution | Kanazawa University |
|---|
Principal Investigator |
KOH Eitetsu Kanazawa University, Graduate School of Medical Science, Associate Professor, 医学系研究科, 助教授 (90283134)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KANEKO Shuichi Kanazawa University, Graduate School of Medical Science, Professor, 医学系研究科, 教授 (60185923)
NAMIKI Mikio Kanazawa University, Graduate School of Medical Science, Professor, 医学系研究科, 教授 (70155985)
MIZOKAMI Atsushi Kanazawa University Hospital, Assistant Professor, 医学部附属病院, 講師 (50248580)
|
|---|
| Project Period (FY) |
2003 – 2004
|
| Keywords | Y chromosome / male infertility / AZF / palindrome / genome / STS / real-time PCR / Spermatogenesis |
|---|
| Research Abstract |
Background :
In 2003, the whole sequence of the Y chromosome has been determined following the completion of the human genome project. As a result, huge identical sequences have been found to be present massive Y-specific repeat called amplicons on the distal side of the euchromatic region of the long arm of the Y chromosome, and these amliconic regions have formed a massive palindrome complex. The palindromes in the AZFc region are consists of a complex of several small segments. These ampliconic sequences pairs are characterised by palindromes showing nearly more than 99.9% identy. Sequence tagged site (STS) markers are now available for detection of the position of palindromes. The AZFc region is comprised completely of amplicons and is particularly susceptible to deletion. The b2/b4 deletion is eliminated the entire AZFc region and cause the spermatogenetic failure. Some papers reported partial deletions within AZFc are present in some infertile male. Conventional PCR is not evaluat
… More
ed a copy number, but, real-time PCR assay that may be adaptable for estimating several identical sequence sites as copies numbers.
The objective of the present study was to evaluate the multiple sites which is identical to sequences using the quantitative detection of copies by real-time fluorescence PCR as a new molecular diagnostic parameter in the genome DNA from patients presenting with non-obstructive azoospermia. Here we apply quantitative real-time PCR as an alternative approach to measure DNA copy number changes at each AZF region of the human Y chromosome.
We selcted the probes such as sY 142 (G38345), sY 254 (G38349), sY 579 (G63909), sY 602 (G34986), sY 627 (G67175), sY 639 (G67162), sY 1054(G6716), sY 1125 (G67164), sY 1190 (G67165), sY 1192 (G67166), sY 1196 (G67167), sY 1197 (G67168), sY 1198 (G67169), sY 1201 (G67170), sY 1206 (G67171). We modified these primers size for real time PCR, so that the PCR products products is amplified approximately 100 base pairs and hybridization probes within each primer and dual-labelled flurogenic probes (hybridization probe) is designed between these selected modified primers. These amplicons and corresponding primers that showed a unique BLAST hit were selected and used in further experiments.
The region of AZFc are present one or more identifical sequences per genome because of their palindromic structures. As more several retetive sequence exist, the possibility of partial deletion within AZFc is present.
We here demonstrated the application of a new, flexible, fast, and precise real-time PCR based estimation the copy number of identical sequences in Y chromosome ampliconic region. Less
|
Research Products
(3 results)
