Vasa Gene Marker for Tuna Germ Cell Detection
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Solution Overview
Problem
Current methods for tuna germ cell transplantation and seedling production are inefficient due to the large size of tuna, requiring significant manpower and costs, and there is a need for a method to distinguish tuna-derived germ cells from those of the recipient fish in surrogate fish techniques.
Innovation Solution
The use of specific Vasa gene sequences and nested PCR combined with restriction enzyme treatment to detect and distinguish tuna-derived germ cells in the genital gland of a recipient fish, allowing for the successful transplantation and growth of tuna germ cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional methods are used for tuna germ cell transplantation, then the process can be performed, but it requires significant manpower and costs due to the large size of tuna
Solution Approach 1:
The patent replaces mechanical/manual handling methods with molecular biological detection methods. Specifically, it uses species-specific marker genes and PCR-based detection systems to identify and track transplanted germ cells, eliminating the need for manual manipulation and visual inspection that would require significant manpower for large tuna.
Solution Approach 2:
The patent introduces molecular markers (species-specific genes or genetic modifications) as intermediaries to facilitate the detection and tracking of transplanted germ cells. These markers serve as detectable signals that allow researchers to monitor germ cell transplantation success without direct mechanical intervention in the large tuna organisms.
2Measurement precision
If traditional detection methods are used, then germ cell transplantation can be monitored, but it cannot reliably distinguish tuna-derived germ cells from recipient fish germ cells
Solution Approach 1:
The patent applies local quality by introducing species-specific molecular markers into the germ cells. These markers are localized to specific genetic regions or expressed as specific proteins that differ between donor tuna and recipient fish, enabling precise differentiation at the molecular level rather than requiring whole-organism or tissue-level analysis.
Solution Approach 2:
The patent uses molecular copying techniques, specifically PCR amplification of species-specific marker genes, to generate detectable signals from the transplanted germ cells. By copying and amplifying unique genetic sequences from the donor tuna, the system can detect and distinguish these cells from recipient fish cells even when present in small numbers within the recipient's gonads.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This method enables reliable detection and differentiation of tuna germ cells, facilitating efficient seedling production and growth by ensuring the successful incorporation and survival of tuna-derived germ cells in the recipient fish.
Implementation Method 1
differential expression of vasa RNA and protein during spermatogenesis and oogenesis
Implementation Method 2
a method for detecting the germ cell of Perciformes fish such as tuna, using such Vasa protein or Vasa gene as a target
Implementation Method 3
The use of specific Vasa gene sequences and nested PCR combined with restriction enzyme treatment to detect and distinguish tuna-derived germ cells
Implementation Method 4
nested PCR combined with restriction enzyme treatment to detect and distinguish tuna-derived germ cells
Data Source
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AI summary
In order to examine whether or not a germ cell derived from a donor fish, which has been transplanted into a recipient fish of a different species by a surrogate fish technique, grows or matures in the gonad of the recipient fish, it is necessary to use, as an indicator, a trait that is specifically expressed in the germ cell and can be used to distinguish the recipient fish from the donor fish. Vasa gene, which is a germ cell-specific gene, is specific to a primordial germ cell and a spermatogonium/an oogonium, and it is not expressed in a somatic cell. In the present invention, the Vasa gene sequences of a tuna, a chub mackerel, a spotted mackerel, an eastern little tuna, and a drumfish are determined, and the expression of such gene is used as a marker for a germ cell. In addition, according to the present invention, it is possible to specifically detect only a tuna Vasa gene in Vasa gene sequences that are highly conserved in fishes, without sequencing. Thus, a tuna-derived germ cell can be reliably and simply identified in the gonad of the recipient fish. As a result, the growth or breeding of tuna can be carried out with good efficiency. Moreover, utilizing the aforementioned findings, even in a case in which not only a tuna but also another Perciformes fish is used as a donor, a germ cell derived from the donor fish can be efficiently detected from the gonad of a recipient fish of a different species.